Abstract
Fibroblasts, the principal cell type of connective tissue, secrete extracellular matrix components during tissue development, homeostasis, repair and disease. Despite this crucial role, the identification and distinction of fibroblasts from other cell types are challenging and laden with caveats. Rapid progress in single-cell transcriptomics now yields detailed molecular portraits of fibroblasts and other cell types in our bodies, which complement and enrich classical histological and immunological descriptions, improve cell class definitions and guide further studies on the functional heterogeneity of cell subtypes and states, origins and fates in physiological and pathological processes. In this review, we summarize and discuss recent advances in the understanding of fibroblast identification and heterogeneity and how they discriminate from other cell types.
Similar content being viewed by others
Introduction
Fibroblasts were first described in 1858 by Rudolf Virchow as cells located in connective tissue1, a term coined already in 1830 by Johannes Peter Müller for the matter that connects and mechanically supports other tissues2. In 1879, Matthias Duvall distinguished fibroblasts from epithelial cells within the mesenchyme of chick embryos, thereby adding to their demarcation3. Progressively, it became clear that fibroblasts reside within all dense and loose fibrous connective tissues. Dense fibrous connective tissue makes up tendons, ligaments and fasciae of the musculoskeletal system. Loose fibrous connective tissue is located at many sites, including visceral organ capsules, beneath the epithelial linings of airways, gastrointestinal and urogenital tracts (laminae propria) and skin (dermis), in central nervous system meninges, and between parenchymal cells in muscle and adipose tissue. Fibroblasts are thus a major cellular component of the diverse fibrous connective tissues that disperse throughout our organs2,4. Alongside endothelial and blood cells, fibroblasts are one of the most common and widespread cell types in our bodies2.
The term fibroblast, used already more than 100 years ago by Ernst Ziegler5 and Alexander Maximow6, deserves a comment, as the “-blast” suffix refers to morphological hallmarks of active protein synthesis. A key research goal of the early 20th century was to identify the cellular source of extracellular matrix (ECM) during tissue repair4,7,8,9. In these studies, the quiescent fibroblasts of uninjured organs were sometimes referred to as fibrocytes, which displayed a small cytoplasm, few ribosomes and condensed chromatin, together suggesting low levels of protein synthesis10. It was assumed that fibrocytes give rise to fibroblasts upon wounding, and conversely that fibroblasts differentiate into fibrocytes during healing10. Perhaps due to limited interest in the quiescent cells, the original meaning of fibrocyte was progressively lost and fibroblast adopted as the common name irrespective of its state of activity. Fibrocyte later reappeared as the name for a circulating monocytes/macrophages-related antigen-presenting cell involved in tissue repair11,12 (for review see ref. 13).
In the early days of cell culturing, fibroblasts turned out to be sturdy and easy to propagate on artificial surfaces such as glass and plastic. They regularly outgrew other cell types and thus became popular for basic studies in cell biology. In the 1950s, Michael Abercrombie and Joan Heaysman established the important concept of cell contact inhibition using fibroblasts14, and Leonard Hayflick showed in 1965 that fibroblasts stop dividing and senesce after a finite number of population doublings in vitro15, thereby disproving the view that cultured cells could be passaged indefinitely (see ref. 16). Fibroblasts were further utilized to identify cellular oncogenes in the 1970s and 1980s17, as well as for the more recent establishment of techniques for induced pluripotency18. Fibroblasts are also commonly used as feeder cells in co-culture experiments to provide cell contacts, ECM and growth factors needed for other cells, such as embryonic stem cells, to thrive19,20.
It is well-established that fibroblasts produce collagens and numerous other proteins that make up the ECM of fibrous connective tissues. In fact, one of the fibroblasts’ archetypical products, collagen type 1, is the most abundant protein in our bodies21. The ECM consists of more than 300 different core proteins and numerous additional bridging proteins and matrix-modifying enzymes, together referred to as the matrisome22. Each tissue has its unique ECM composition, and it was early recognized, based both on morphological and molecular criteria, including gene expression profiling, that fibroblasts from various anatomical locations differ23,24. In addition to ECM production, fibroblasts play a role in wound contraction. In 1971, Giulio Gabbiani and colleagues reported that fibroblasts respond to tissue injury by assuming a contractile, myofibroblast, phenotype25. Similar cells are implicated in fibrosis, which may be defined as unresolved repair after tissue damage (for review, see ref. 26). Besides fibrosis, fibroblasts are important also for other disease processes, including cancer where they make up a variable proportion of the tumor stroma and deposit ECM of various molecular compositions and secrete different cocktails of growth factors and cytokines that influence tumor growth and metastasis27,28. Secretion of soluble extracellular signaling molecules is also of profound physiological importance during development, where fibroblasts provide instructive paracrine signals during organogenesis and thus take part in reciprocal cell-cell signaling required for the differentiation of other—often epithelial—cell types, for example during epidermal, oral, intestinal and kidney development29,30,31,32,33,34. Finally, fibroblasts are known to hold positional memory in the body, notably by expression of specific sets of Hox transcription factors in various sectors along the craniocaudal axis of the vertebrate embryo35,36.
To better understand the various roles of fibroblasts, it is important to gain insights into how heterogenous they are, as well as the functional consequences emanating from fibroblast heterogeneity. At least in part, research into fibroblast heterogeneity has however been hampered by ambiguous and diversified cell classification with regard to, for example, lineage, morphology, location or growth characteristics. An important step towards a deeper and more detailed characterization of fibroblasts can however now be taken thanks to single-cell transcriptomics, which complements the detailed anatomical and morphological knowledge with insights into the molecular profiles of fibroblasts at unprecedented depth and scale.
Evolution from morphological to molecular characterization of fibroblasts
In normal adult tissues, fibroblasts appear spindle- or stellate-shaped with an oval nucleus and a distinct endoplasmic reticulum (ER)3,4 (for review see ref. 27). While these features are helpful to localize the cells anatomically, they provide few clues to origin, molecular composition, heterogeneity and relationship to other cell types. Although not uniquely expressed on fibroblast, certain proteins, such as vimentin (VIM (protein), Vim (gene/mRNA mouse)) and fibroblast specific protein 1 (FSP1, S100a4), have served as useful markers to identify fibroblasts by immunohistochemical techniques37,38. With the advent of technologies that allow the gene expression patterns of individual cells to be resolved at substantial depth, i.e., single-cell RNA-sequencing (scRNA-seq) (see Box 1 for a detailed description of the technology and potential caveats and pitfalls), an additional important step could be taken in the quest to better characterize fibroblasts along with all other cell types that make up our organs. Hence, the traditional morphological classification of cell types is now being complemented by molecular classifications, foremost based on gene expression.
Following from the first cDNA microarray-based study of fibroblast heterogeneity24, transcriptional analysis of fibroblast was conducted using bulk mRNA isolates obtained from multiple cells39,40,41. This work provided molecular information at the organ level and initial insights into inter-organ transcriptional heterogeneity of fibroblasts. From these and other studies, certain molecular markers were proposed to distinguish fibroblasts from other cell types, including VIM, platelet derived growth factor receptor-alpha (PDGFRA, Pdgfra), fibroblast activation protein-alpha (FAP, Fap), FSP1; and CD90 (Thy1). Subsequently, scRNA-seq datasets have been obtained from fibroblasts from essentially all major organs in the mouse and human (see Supplementary data 1 for a compilation of select scRNA-seq studies covering fibroblasts). When fibroblasts were collected and sequenced as part of broader atlas projects, they were occasionally called by other names, such as stromal cells, mesenchymal stem cells, myofibroblasts42 and unknown mesenchymal cells43, illustrating the prevailing ambiguities regarding markers and cell nomenclature. Some studies have been devoted more specifically to fibroblasts and their involvement in cancer, fibrotic diseases44,45 or development46. We have ourselves compared fibroblasts across healthy adult organs in the mouse47, whereas others have reported multi-organ comparisons between mice and humans48.
What have we learned from these scRNA-seq studies thus far? As a brief account (more specific examples and discussion are provided below), one lesson is that no single marker appears capable to discriminate all fibroblasts from all other cell types across organs; combinations of markers are needed. A second lesson is that organotypic fibroblast heterogeneity is profound47, although certain marker combinations may point to fibroblast subtypes that cross organ boundaries. Examples include peptidase inhibitor 16 (Pi16) and collagen type 15 alpha-1 (Col15a1) expression, which define subtypes present in several organs48, a Wif1+Comp1+ fibroblast subtype observed in heart valve as well as skeletal muscle perimysial fibroblasts, and two fibroblast subtypes present in both colon and bladder, defined by Tnc+Cd34− and Tnc-Cd34+ expression, respectively47. Thirdly, insights into the specialized physiological functions of various fibroblast subtypes are emerging, for example in human skin, where some fibroblast subtypes appear specialized for ECM production and others for immunological and antimicrobial activities49, or in mouse skin, where fibroblast subtypes have been identified that participate in fibroblast growth factor (FGF) and Wnt signaling with hair follicle epithelial stem cells during cycling of the hair follicle50. In the mouse intestine, specific intestinal fibroblast subtypes are involved in Wnt signaling that regulates intestinal epithelial stem cell differentiation32,47,51,52. Fourthly, insights into the specialized pathological functions of various fibroblast subtypes are emerging. For example, a rare intestinal fibroblast subtype that converts arachidonic acid into prostaglandin E2 appears to play a role in tumorigenesis in the mouse33. Fibroblasts in skeletal muscle express specific cytokines and proinflammatory factors, likely aiding in regeneration of myofibers following injury53. A fifth lesson relates to species similarities and differences, including how similar human and mouse fibroblasts are, which is of relevance for human translation of mouse data. While the number of studies comparing mouse and human fibroblasts “side-by-side” are still few48,54,55,56,57,58, analogous fibroblast subtypes and similar gene activation programs exist in mouse and human cancer-associated fibroblasts (CAFs)48. There also appears to be molecular similarities between perivascular brain fibroblasts in humans and mice59,60,61,62, as well as between human and mouse lung fibrosis-inducing CTHRC1+ fibroblast subtypes63,64. Notwithstanding these examples, more research will be required to precisely define the degree of fibroblast conservation between the humans and mice.
Given the current pace of acquisition of fibroblast scRNA-seq data, we will undoubtedly see rapid progress, resulting in more fine-grained views of fibroblast heterogeneity, as well as refinement of transcriptomic signatures for cell type demarcations, such as proposed pan-fibroblast signatures defined by Pdgfra, Dpt combined with Pi16 or Col15a148, or a 90-gene signature that demarcates fibroblasts from vascular mural cells47. Progress will also be aided by improvements on the bioinformatics side (see Box 1 regarding technological considerations for scRNA-seq studies). Going forward, it is important that novel fibroblast subtypes are rigorously characterized, not only bioinformatically through clustering algorithms, which can be somewhat arbitrary and should be seen as a framework for detailed analysis rather than final conclusions about cell types, but also anatomically using carefully annotated RNA and protein markers. Ideally, marker combinations that readily and reliably distinguish fibroblasts from other cell types in specific organs of interest should be identified and used.
Fibroblast heterogeneity within and between organs
As mentioned above, scRNA-seq data now allow fibroblast heterogeneity and relationship with as well as demarcation to other cell types to be decoded with unprecedented speed and precision. When discussing progress in these areas, we were inspired by the “The Ancestor’s Tale” by Richard Dawkins and Yan Wong, in which the authors describe our evolutionary history from the perspective of a human first discussing with its closest relatives, i.e., the extinct hominins, and then proceeding gradually to evolutionarily more distant relatives, ending with Archaea. In the same vein, we first discuss fibroblast heterogeneity within and between organs and then proceed to discuss relationships of fibroblasts to other connective tissue and vasculature cell types with whom fibroblasts share functional similarities and possible molecular relationships.
Intra-organ fibroblast heterogeneity
Fibroblast heterogeneity spans species, organ and developmental stage boundaries. It has long been recognized that different fibroblast populations reside simultaneously in the same organ (for review see refs. 65,66). For example, several studies have advanced our understanding of fibroblast subtypes in the skin (Fig. 1). In mice, fibroblasts residing in the upper (papillar) layer express alpha-8 integrin (Itga8) and dipeptidyl peptidase 4 (CD26, Dpp4), whereas fibroblasts in the lower (reticular) layer express podoplanin (Pdpn) and delta-like non-canonical Notch ligand 1 (Dlk1)67. One study identified four fibroblast subtypes in the adult mouse skin (FIB1-4), which based on marker expression (Dcn, Gpx3, Sparc, and Plac8, respectively), were found to occupy three distinct anatomical localization (dermis, hypodermis and adventitia)50. Two of the subtypes (FIB1 and 2), likely representing different states of the same fibroblast subtype, respond transcriptionally to different cell cycle stages in the hair follicle50. In humans, one study suggests that skin fibroblasts segregate into six transcriptionally distinct cell clusters, of which one (expressing DPP4) is the main ECM-producing cell type49. In a meta-analysis of human skin fibroblast transcriptomes, 10 subtypes formed three major groups (A-C, marked by MMP2, IGFBP7 and SFRP1 expression, respectively)68, of which group A appears specialized for ECM production and group B in immune surveillance68. In the mouse heart, two major fibroblast subtypes, presumably derived from endo- and epicardium, were identified and shown to express Lamb1 and Dkk3, respectively69. Heart fibroblasts also express Csf1, Vegfa, Igf1 and Fgf2, indicating active paracrine signaling to other cell types in the heart69. The human adult heart was found to contain seven fibroblast subtypes, two of which show enrichment in atria and ventricles and can be distinguished by expression of SCN7A and CFH, respectively, and two (called FB4 and FB5) which appear specialized in responding to transforming growth factor-β signaling and in ECM remodeling, respectively70. In the mouse lung, fibroblast subtypes have been characterized, one of which (a PDGFRa+Axin2+ subtype) responds by differentiating into myofibroblasts following injury71,72. An analysis of the human lung revealed five fibroblast subtypes, one of which (CTHRC1+) promotes lung fibrosis in COVID-19 patients64. Interestingly, a similar Cthrc1+ fibroblast subtype drives lung fibrosis in the mouse63. Two fibroblast subtypes with different injury-response features were identified in the synovial tissue of joints in the mouse: an immune effector subtype expressing fibroblast activation protein-α (FAPα)+ as well as THY1+ and a tissue destructive FAPα+THY1− subtype73. Depletion of the FAPα+ fibroblasts reduced bone erosion and inflammation in a mouse arthritis model73. Another study identified NOTCH3 activity as a driver of inflammation in THY1+ synovial fibroblasts44. Intra-organ fibroblast heterogeneity is not confined to mammals but has also been noted in zebrafish where different pdgfra-positive fibroblast subpopulations take part in lymphangiogenesis46, or in the axolotl where several fibroblast populations were found amongst blastema-cells during limb regeneration74,75.
In addition to the distinction between fibroblast entities by the presence/absence of specific singular markers, intra-organ fibroblast heterogeneity may also reflect cell zonation, which is defined as gradual cellular phenotypic transitions along an anatomical or functional axis. For example, zonation of hepatocytes and endothelial cells occurs along the porto-central axis in the liver lobule76,77 and endothelial and mural cell zonation is observed along the arterial-venous axis in brain vasculature60. There are anatomical and physiological reasons to believe that fibroblast zonation (in addition to the Hox transcription code, discussed above) would make sense, for example along the muscle-tendon axis or along the crypt-villus and crypt-surface axes of the small and large intestine32,34. By identifying genes with gradient or nested transcription profiles across adjacent tissue areas/volumes, fibroblast zonation will potentially be uncovered and shown to be instrumental for generation and maintenance of proper tissue architecture in multiple organs.
Inter-organ fibroblast heterogeneity
The first genome-wide transcriptional information on inter-organ fibroblast heterogeneity was provided through cDNA microarray analysis of cultures of human skin fibroblasts established from different ages and anatomical locations24. Interestingly, despite the concern that in vitro culturing may have influenced gene expression patterns, these fibroblasts retained a certain degree of transcriptional memory of their origin, including HOX gene expression patterns24. In a cross-comparison of mouse fibroblasts isolated directly from different muscular organs and analyzed by scRNA-seq, it was evident that differential expression of genes related to the ECM—the matrisome—is a major feature of inter-organ fibroblast transcriptomic heterogeneity47 (Fig. 2). This information is consistent with previous evidence that fibroblasts at different anatomical locations and at different ages tailor a unique composition of the ECM24. An important topic for further analysis is to understand the genetic and/or epigenetic underpinnings of how fibroblasts of different organs regulate their gene expression in order to achieve spatial/temporal-specific ECM profiles.
The relationship between fibroblasts and other cell types in connective tissue and vasculature
Specialized connective tissue cell types
Several connective tissues harbor highly specialized cell types: adipocytes in fat, chondrocytes in cartilage, and osteocytes/blasts in bone. There are both in vivo and in vitro data suggesting that these cell types are lineage-related and share a common mesenchymal and fibroblast-like origin, and it is therefore of interest to explore how related the differentiated cells are to each other and to fibroblasts at the molecular level. Adipocytes are designed to store, release, and burn lipids at different relative rates depending on their subtype (white, brown, and beige/brite). Several scRNA-seq studies of adipocytes have been conducted, although adult white adipocytes are challenging to handle due to their large size and fragility (for review see ref. 78). From available data, it is apparent that the transcriptomes of differentiated fibroblasts and adipocytes are readily distinguishable by the adipocyte transcripts that encode proteins involved in lipid handling. The resemblance between the adipocyte precursor cell (preadipocyte) and fibroblasts is however close, although relationships between preadipocytes and pericytes, discussed below, have also been proposed (see ref. 79 and references therein). Chondrocytes are morphologically highly similar to fibroblasts, but express several unique proteins, notably a number of ECM proteins contained within cartilage, including collagen type 280 (for review see ref. 81). Beyond the highly specific markers of adipocytes, chondrocytes and osteoblasts, further analyses are needed to define transcriptional similarities and differences between these cells, their precursors, and fibroblasts.
Vascular mural cells
Vasculature courses through and is sometimes even counted as a component of connective tissue. Blood vessels consist of an inner tubing of endothelial cells surrounded by mural cells. The latter are either vascular smooth muscle cells (VSMCs), located in larger vessels (arteries, arterioles and veins), or pericytes, which coat capillaries and some venules and are transcriptomically related to venous VMSCs60. VSMCs contain a contractile machinery involving several proteins, among them alpha-smooth muscle actin (ASMA, Acta2), which is a highly expressed and often-used smooth muscle cell marker. Pericytes probably have multiple and organ-specific functions, for example in the brain where they are essential for the maturation and maintenance of the blood-brain barrier82,83. Molecularly, pericytes can be distinguished from VSMCs by for example abundant expression of potassium inwardly rectifying channel subfamily J member 8 (Kcnj8) and absent or very low (relative to VSMCs) expression of transcripts encoding the smooth muscle contractile machinery60. Fibroblasts reside on the outside of the mural coat in many vessel types. In the largest arteries and veins, fibroblasts form an anatomically discernable vascular layer, the adventitia. Pericytes have a morphology somewhat resembling that of fibroblasts, and it has been proposed that pericytes are in fact specialized perivascular fibroblasts84. Live imaging in zebrafish embryos indeed suggest that mural cells derive from perivascular fibroblast-like cells85. ScRNA-seq analysis of pericytes from several organs, including brain, lung, skeletal muscle, colon, bladder and heart has illustrated their relationship to fibroblasts47,60. From these studies, it seems clear that a distinct demarcation between fibroblast and pericyte transcriptomes can be made, including differences in ECM-related transcription profiles. For example, pericytes express specific ECM-binding proteins, such as basal cell adhesion molecule (BCAM, Bcam), but generally low levels of collagens in comparison to fibroblasts, with the exception of collagen type 4 (Col4a1, Col4a2), which are components of the microvascular basement membrane to which the pericytes contribute47. The molecular demarcation between VSMCs and fibroblasts also seems distinct, with several genes uniquely expressed in one or the other cell type. Regarding ECM production, VSMCs express low amounts of collagens, similar to pericytes, but high amounts of elastin (Eln), and certain other ECM proteins of presumed relevance for the resilience of the arterial wall47. Genes encoding proteins of the smooth muscle contractile machinery, such as Acta2, myosin heavy chain 11 (Myh11), transgelin (Tagln), calponin 1 (Cnn1), leiomodin 1 (Lmod1) and regulator of calcineurin 2 (Rcan2), are highly expressed by VSMCs, distinguishing them from hitherto analyzed quiescent fibroblasts, although these genes may be re-expressed (albeit at lower levels than in VSMCs) when fibroblasts become activated into myofibroblasts during pathological conditions or in normal fibroblasts located in organs with high deformation capacity, such as the urinary bladder47,57.
Cell types with features of both fibroblasts and pericytes
While in most cases fibroblasts and pericytes can be readily distinguished by their distinct gene expression profiles, certain cell types appear to exhibit characteristics of both. In the kidney glomerulus (Fig. 3A), mesangial cells display intermediate features between those of pericytes and fibroblasts, VSMC and myofibroblasts86. Mesangial cells have indeed been proposed to be a specialized type of pericytes87,88, a notion supported by their lineage-dependence on PDGFB-PDGFRB signaling89,90,91, which is shared with other mural cell lineages. Other reports however claim that mesangial cells are distinct from pericytes92. Mesangial cells lack the typical cellular morphology and vascular basement membrane embedment of other capillary pericytes, and the fact that they deposit a rich ECM (called mesangial matrix) may be more in line with a fibroblastic than pericytic nature. However, no other pericyte-like cells are present within the glomerulus, and if a generic pericyte function were needed there, it would have to be provided by the mesangial cells. Currently, several kidney scRNA-seq studies have claimed to identify (and hence molecularly characterize) mesangial cells, but links between cellular heterogeneity to precise anatomical location were lacking93,94,95. However, one study succeeded in distinguishing between intra- and extraglomerular mesangial cells through their unique scRNA-seq profile and in situ analysis of relevant markers58. Notably, the intraglomerular mesangial cells expressed desmin (Des) and Acta2, i.e., markers of mural cells, but also Pdgfra and Pi16, which are fibroblast markers typically absent in mural cells58.
Hepatic stellate cells (HSCs) were originally discovered in the 1870s by Karl Wilhelm von Kupffer, who called them Sternzellen (star cells)96 and portrayed them as a specialized type of pericyte by virtue of their anatomical position along the sinusoidal capillaries and in the perisinusoidal space (the space of Disse) between the endothelial cells and hepatocytes97,98,99,100 (Fig. 3B). Like pericytes in other organs, the HSCs contact capillary endothelial cells, but unlike both typical pericytes and glomerular mesangial cells, HSCs do not depend on PDGFB-PDGFRB signaling for their embryonic development101, possibly suggesting a non-mural identity. HSCs were recently characterized by scRNA-seq and found to be heterogeneous across the healthy liver lobule, with distinct molecular markers expressed at the periportal and central locations, respectively102. Similar to mesangial cells, HSCs appear to express markers of both mural cells (Des) and fibroblasts (Pdgfra), suggesting an identity intermediate between these two cell types.
Annotation of ambiguous cell types
The emerging transcriptomic profiles of fibroblasts and the other cell types discussed above can be useful to shed light on the nature of recently identified cell types, for which annotation has been problematic, or remains controversial. A novel type of pericyte involved in spinal cord repair has been reported103,104. While these cells share some markers with brain pericytes and locate in the proximity of blood vessels, other markers may argue that these cells instead are perivascular fibroblasts105, a notion receiving support from scRNA-seq data60. Lineage-tracing studies likewise suggest a fibroblast origin of cells forming spinal cord fibrotic scars106, but a final conclusion will have to await a more extensive analysis of all cell types present in the spinal cord vasculature. Kidney fibroblasts were recently annotated in a kidney scRNA-seq study based on the expression of a singular marker, FSP1 (S100a4)107. However, cross-organ comparison provides little support for these cells being fibroblasts, but there is instead ample evidence for similarity with macrophages47,108, which, along with smooth muscle cells, also express FSP1. Another study instead annotated kidney fibroblasts in both human and mouse as positive for PDGFRA, PDGFRB, maternally expressed 3 (MEG3), Scavenger Receptor Class A Member 5 (SCARA5) and numerous collagens109, which are markers found also in fibroblasts from several other organs47. Further studies are thus needed to establish the identity and possible heterogeneity of kidney fibroblasts. Some of the annotations in the large-scale analysis presented in Tabula Muris42 may also need to be revised, notably some cell populations defined as myofibroblasts should rather be referred to as mural cells (pericytes and/or VSMCs). A novel lung cell type characterized by high PDGFRB expression and expansion in response to experimental lung injury was recently identified110. The authors posed the question whether these cells were pericytes, a notion supported by the extensive transcriptomic similarity to cells independently annotated as lung pericytes60. These examples illustrate how the improved understanding of fibroblasts, pericytes and other vascular or connective tissue cell types may assist in fine-tuning annotations of cell types from various datasets.
Lineage relationships between fibroblasts and other cell types
The recent advances in transcriptomics can be productively interfaced with insights from lineage-tracing analysis to better understand fibroblast lineage relationships. Connective tissue is largely derived from mesoderm that via epithelial-to-mesenchymal transition (EMT) of the primitive epithelium generates embryonic mesenchyme, which in turns forms most connective tissues in the adult organism, including fibroblasts65,111 (Fig. 4). The vascular mural cells of most trunk tissues likewise have a mesodermal origin, arguing that a common progenitor for fibroblasts and vascular mural cells can be found somewhere in the mesodermal lineage65, a view supported by lineage-tracing analysis112. Fibroblasts and mural cells in the head are however in part derived from a different origin, the neuroectoderm (neural crest)84, and it will be interesting to learn to what extent transcriptomic differences between fibroblasts and mural cells in the head versus organs in the trunk reflect these different cellular origins. To establish fibroblast transcriptomes from different developmental stages will also be important to understand how different fibroblast subtypes dynamically change during development.
In a discussion on connective tissue lineage and possible stem/progenitor populations, it is difficult to avoid touching upon mesenchymal stem cells (MCS; a.k.a. mesenchymal stromal cells). MSCs were first mentioned in the 1960s in conjunction with bone marrow transplantation and explant studies113,114, and cells with the potential of differentiating into multiple connective tissue phenotypes, including adipose, cartilage and bone, were subsequently established in vitro from multiple tissue sources (for review see ref. 115). There has however been a lack of rigorous criteria to define MSCs116 and although efforts are made to define a set of commonly agreed markers115,117, there is still a somewhat bewildering literature regarding MSC cell lineage and differentiation capabilities. It should also be considered that most MSC differentiation experiments have been conducted in vitro, which makes it difficult to yet establish their true existence in vivo, both regarding abundance and locations, and to what extent they contribute to repair and remodeling of connective tissue, including fibroblasts. Several studies propose that pericytes may be in vivo equivalents to MSCs based on that MSCs can be established from explanted vascular fragments and by using a limited set of markers118,119,120. A strong similarity between MCSs and fibroblasts has however also been proposed121. It is increasingly clear that fibroblasts coat also small caliber vessels without a discernable adventitia and that fibroblasts and mural cells share many markers including the ones used for MSCs47,60. In sum, MCSs have been extracted from various tissues, but their true in vivo origin, i.e., whether they exist as a bona fide resident stem/progenitor cell in the tissue or derive through dedifferentiation of a tissue-resident fibroblast, pericyte or other cells, remains to be worked out.
Lineage-tracing in the mouse has shed new light on relationships between fibroblast subtypes and between fibroblasts and other cell types, although there are still some technical caveats and pitfalls with the technology, as discussed in Box 2. In skin, progenitors for both papillar and reticular fibroblasts, as well as for the papillar and reticular lineages individually, have been identified by the use of Pdgfra-CRE, Blimp1-CRE and Dlk1-CRE drivers, respectively67 (for review see ref. 122). Skin fibrosis has been lineage traced to an En1+ fibroblast population123. During wound healing, cells descending from a Hic1+ mesenchymal progenitor population generate extrafollicular fibroblasts124, and Hic1 marks mesenchymal progenitors with functions in regeneration also in skeletal muscle53 and heart125. Interestingly, these cells express a number of pericyte markers (e.g., Rgs5, Mcam, Notch3 and Kcnj8), possibly suggesting a lineage relationship between pericytes and regenerating fibroblasts53. In heart, mesenchymal progenitors yield Pdgfra+Sca1− fibroblasts upon cardiac injury, and the transition towards the fibroblast state is regulated by Hic1125. Lineage-tracing of a Pdgfra+Ly6a+(Sca1) mesenchymal stromal progenitor cell in skeletal muscle indicates that it can give rise to both fibroblasts and adipocytes126,127 and be diverted towards an osteogenic fate by bone morphogenetic protein stimulation128. This may suggest a common mesenchymal progenitor for fibroblasts, adipocytes and osteoblasts.
Fibroblasts and pathological conditions
Fibroblasts have been strongly implicated as the major fibrogenic cell type in pathological tissue fibrosis. 50 years ago, it was demonstrated that fibroblasts convert to myofibroblasts expressing certain smooth muscle cell markers, notably Acta2, when activated by tissue injury or stress25. While this transition is part of a normal wound-healing process that resolves over time, it can also result in persistent fibrosis, a severe medical condition with impaired function of organs, including the kidney, lung, liver and heart. A hallmark of fibrosis is the accumulation of myofibroblasts and excessive ECM deposition resulting in abnormal fibrous connective tissue (for review see refs. 27,129). While many studies point to fibroblasts as the source of the myofibroblast, the situation may not be that simple. Alternative suggested origins of myofibroblasts include endothelial cells, macrophages, mesothelial cells (the squamous epithelial lining of body cavities and their organs), pericytes, HSCs and MCSs. Summarizing the extensive body of original papers regarding the potential diverse origins of myofibroblasts goes beyond the scope of this review, and we refer interested readers to other literature on the topic66,129,130. Lineage-tracing has identified a Gli1+ mesenchymal progenitor population, which generates myofibroblasts and drives fibrosis in a sonic hedgehog (Shh)-dependent manner in kidney, lung, liver, heart and bone marrow131,132,133. The localization of the Gli1+ mesenchymal cells close to endothelial cells in the perivascular niche and their expression of Cspg4 (NG2) may suggest a relationship to pericytes131. The partially conflicting propositions for the various cellular origins of myofibroblasts rest on lineage-tracing experiments in mice and may to some extent reflect methodological pitfalls, including leakiness in inducible CRE/loxP systems134 or aberrant expression of CRE recombinase (Box 2). It however remains a possibility that there is a multi-cell type (including non-fibroblastic) origin generating functionally distinct subpopulations of myofibroblasts. Heterogeneity of fibroblast origin and differentiation during tissue fibrosis is indeed supported by data from heart, lung and kidney109,135,136. Further careful assessment of transcriptomic data from resting fibroblasts and other normal cell types, comparisons with myofibroblast transcriptomes, and the analysis of transcriptional profile changes during myofibroblast generation109 should bring clarity to both origin and heterogeneity amongst pathological myofibroblasts.
In cancer research, it is increasingly realized that it is not only the properties of the genetically abnormal tumor cells that dictates the cancer’s severity, growth potential and ability to metastasize; the outcome is also influenced by the genetically normal stroma cells that populate all solid tumors at variable abundance. Among the latter, CAFs constitute an important cell population (for a recent consensus statement paper on CAFs, see ref. 137). Together with their produced matrix, CAFs often make up a significant volume of carcinomas, and they influence tumor growth and metastasis27,28. Similar to the situation with myofibroblasts and fibrosis, our knowledge about the origin and molecular characteristics of CAFs is still limited—they are in part “negatively” characterized by not expressing markers for other well-annotated cell types and not carrying the chromosomal abnormalities observed in the neoplastic tumor cells. To gain insights into the origin of CAFs is complicated, particularly in humans, and relies on inference from studies of earlier stages of cancer (hyperplasia and adenomas), and based on such studies, normal tissue fibroblasts have been implicated as a founding cell type for CAFs (for review see ref. 138). Lineage-tracing experiments in mice have resulted in different views on the cell type of origin for CAFs137, but the conclusions may suffer from problems with CRE expression specificity (Box 2), as discussed above for myofibroblasts. Cell transplantation experiments provide an additional source of information regarding the cellular origin of CAFs. In such experiments, asking whether a particular transplanted cell type ends up as CAFs in the tumor stroma, both MSCs and adipocytes appear to be able to generate CAFs in xenograft mouse tumor models139,140,141,142,143. As for myofibroblasts, it is an emerging view that distinct subpopulations of CAFs can be identified in a particular tumor stroma45, possibly indicating a multi-cell type origin of CAFs.
Conclusions and future outlooks
At present, our understanding of fibroblasts is rapidly improving, and large-scale transcriptomic analyses at single-cell resolution allow us to better define fibroblasts, their heterogeneity and relationship to other types and classes of cells. While this undoubtedly is transforming our current understanding, it is important to also consider that there still is room for improvement regarding the current transcriptomic and transgenic technologies (see Box 1 and Box 2 for examples). Transcriptomic technologies indeed rapidly improve and become more affordable, which will lead to an acceleration in the accumulation of gene expression information with increasing granularity. Similarly, novel lineage-tracing methodologies are at the horizon, which rely less on promoter specificity but more on clonal evolution based on naturally occurring somatic mutations in cells144,145 or the introduction of traceable genetic markers146. Improved bioinformatic algorithms to identify cell state transitions and advances in spatial transcriptomics will likewise help deciphering lineage relationships and the exact anatomic location of fibroblast subsets147. Such studies may shed new light on lineage relationship and provide insights into whether differentiation yields only divergent fibroblast subsets from a founding cell population or whether there may also be convergence from different lineages towards a similar molecular phenotype, for example when fibroblasts of different origins experience similar physiological conditions and challenges.
Undoubtedly, gene expression signatures that define and demarcate fibroblasts will be refined and reveal further intra- and inter-organ fibroblast heterogeneity, advancing our functional understanding of fibroblasts and its related cell types. While this progress in principle is applaudable, it should be remembered that transcriptomic information, even when available with perfect fidelity for single cells, is not the sole basis of cellular heterogeneity. Cellular protein levels may deviate from the corresponding mRNA levels, and posttranslational protein modifications provide an additional level of molecular complexity and heterogeneity. Additionally, lipids, sugars and low molecular weight metabolites and solutes are likely to contribute to cellular heterogeneity independent of transcriptional control. Whereas single-cell analysis of other molecules than DNA and RNA currently has limited depth, we may expect technological advances also in these areas, and therefore our view and understanding of cellular (including fibroblast) heterogeneity will continue to evolve. Secondly, while the wealth of single-cell transcriptomic data can inspire efforts to define an expanding number of molecularly distinct fibroblast subpopulations, it must all the time be asked to what extent the molecular differences are physiologically meaningful; some of the identified cell “subtypes” may reflect states on a developmental trajectory or responses to stress or injury rather than stable functional specializations at steady state. To tease out specific physiological functions for steady state differences in fibroblasts will undoubtedly take longer, but it remains an important future research avenue and is increasingly feasible. It is possible that the fibroblast cell class in due time may be subdivided into functionally meaningful fibroblast subtypes, some of which execute organ-specific physiological functions, while other subtypes may transcend organ boundaries and carry out functions that are not organ-specific, such as the involvement in immune responses or wound healing. The latter may lead to a terminology similar to the one established for T-cells, for which we now recognize a set of functionally distinct subtypes, including T-regs, CD8+ T-cells, CD4+ T-cells and double-negative T-cells. It will be interesting to see how these endeavors will pan out, but it is certainly likely that the fibroblast will be a well-defined and annotated class of cells for which heterogeneities across and within organs as well as developmental and pathological states are understood in a foreseeable future.
References
Virchow, R. Die Cellularpathologie in Ihrer Begruendung auf Physiologische und Pathologische Gewebelehre (Hirschwald A, Berlin, Germany, 1858).
Bloom, W. & Fawcett, DW. A Textbook of Histology. (CRC Press, 1998).
Duvall, M. Atlas d’Embryologiee. (ed. Masson, G) (Libraire de l’académie de médecine, Paris, France, 1879).
Movat, H. Z. & Fernando, N. V. P. The Fine Structure of Connective Tissue I. The Fibroblast. Exp. Mol. Pathol. 1, 509–534 (1962).
Ziegler, E. General Pathology. (William Wood and Company, 1896).
Maximow, A. Über die Zellformen des lockeren Bindegewebes. Arch. f.ür. mikroskopische Anat. 67, 680–757 (1905).
Lewis, M. Development of connective-tissue fibers in tissue cultures of chick embryos (Carnegie Institution of Washington, Washington, 1917).
Stearns, M. L. Studies on the development of connective tissue in transparent chambers in the rabbit’s ear. I. Am. J. Anat. 66, 133–176 (1940).
Stearns, M. L. Studies on the development of connective tissue in transparent chambers in the rabbit’s ear. II. Am. J. Anat. 67, 55–97 (1940).
Porter, K. R. Cell fine structure and biosynthesis of intracellular macromolecules. Biophysical J. 4, 167–201 (1964).
Chesney, J. & Bucala, R. Peripheral blood fibrocytes: novel fibroblast-like cells that present antigen and mediate tissue repair. Biochemical Soc. Trans. 25, 520–525 (1997).
Bucala, R., Spiegel, L. A., Chesney, J., Hogan, M. & Cerami, A. Circulating fibrocytes define a new leukocyte subpopulation that mediates tissue repair. Mol. Med. 1, 71–81 (1994).
Chong, S. G., Sato, S., Kolb, M. & Gauldie, J. Fibrocytes and fibroblasts—Where are we now. Int. J. Biochem. Cell Biol. 116, 105595 (2019).
Abercrombie, M. & Heaysman, J. E. M. Observations on the social behaviour of cells in tissue culture. I. Speed of movement of chick heart fibroblasts in relation to their mutual contacts. Exp. Cell Res. 5, 111–131 (1953).
Hayflick, L. The limited in vitro lifetime of human diploid cell strains. Exp. Cell Res. 37, 614–636 (1965).
Shay, J. W. & Wright, W. E. Hayflick, his limit, and cellular ageing. Nat. Rev. Mol. Cell Biol. 1, 72–76 (2000).
Shih, C., Padhy, L. C., Murray, M. & Weinberg, R. A. Transforming genes of carcinomas and neuroblastomas introduced into mouse fibroblasts. Nature 290, 261–264 (1981).
Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–76 (2006).
Rheinwald, J. G. & Green, H. Serial cultivation of strains of human epidermal keratinocytes: the formation keratinizing colonies from single cells. Cell 6, 331–343 (1975).
Evans, M. J. & Kaufman, M. H. Establishment in culture of pluripotential cells from mouse embryos. Nature 292, 154–156 (1981).
Karsdal, M. Biochemistry of collagens, laminins and elastin. (Academic Press, 2019). https://doi.org/10.1016/C2018-0-00074-2.
Hynes, R. O. & Naba, A. Overview of the matrisome-An inventory of extracellular matrix constituents and functions. Cold Spring Harb. Perspect. Biol. 4, 1–16 (2012).
Schor, S. L. & Schor, A. M. Clonal heterogeneity in fibroblast phenotype: Implications for the control of epithelial-mesenchymal interactions. BioEssays 7, 200–204 (1987).
Chang, H. Y. et al. Diversity, topographic differentiation, and positional memory in human fibroblasts. Proc. Natl Acad. Sci. USA 99, 12877–12882 (2002).
Gabbiani, G., Ryan, G. B. & Majne, G. Presence of modified fibroblasts in granulation tissue and their possible role in wound contraction. Experientia 27, 549–550 (1971).
Pakshir, P. et al. The myofibroblast at a glance. J. Cell Sci. 133, jcs227900 (2020).
Kalluri, R. The biology and function of fibroblasts in cancer. Nat. Rev. Cancer 16, 582–598 (2016).
Pietras, K. & Östman, A. Hallmarks of cancer: Interactions with the tumor stroma. Exp. Cell Res. 316, 1324–1331 (2010).
el Ghalbzouri, A., Lamme, E. & Ponec, M. Crucial role of fibroblasts in regulating epidermal morphogenesis. Cell Tissue Res. 310, 189–199 (2002).
Costea, D. E., Johannessen, A. C. & Vintermyr, O. K. Fibroblast control on epithelial differentiation is gradually lost during in vitro tumor progression. Differentiation 73, 134–141 (2005).
Liu, Y. Cellular and molecular mechanisms of renal fibrosis. Nat. Rev. Nephrol. 7, 684–696 (2011).
Degirmenci, B., Valenta, T., Dimitrieva, S., Hausmann, G. & Basler, K. GLI1-expressing mesenchymal cells form the essential Wnt-secreting niche for colon stem cells. Nature 558, 449–453 (2018).
Roulis, M. et al. Paracrine orchestration of intestinal tumorigenesis by a mesenchymal niche. Nature 580, 524–529 (2020).
Shoshkes-Carmel, M. et al. Subepithelial telocytes are an important source of Wnts that supports intestinal crypts. Nature 557, 242–246 (2018).
Alexander, T., Nolte, C. & Krumlauf, R. Hox Genes and Segmentation of the Hindbrain and Axial Skeleton. Annu. Rev. Cell Dev. Biol. 25, 431–456 (2009).
Rinn, J. L., Bondre, C., Gladstone, H. B., Brown, P. O. & Chang, H. Y. Anatomic demarcation by positional variation in fibroblast gene expression programs. PLoS Genet. 2, e119 (2006).
Dulbecco, R., Allen, R., Okada, S. & Bowman, M. Functional changes of intermediate filaments in fibroblastic cells revealed by a monclonal antibody. Proc. Natl Acad. Sci. USA 80, 1915–1918 (1983).
Strutz, F. et al. Identification and characterization of a fibroblast marker: FSP1. J. Cell Biol. 130, 393–405 (1995).
Heruth, D. P., Gibson, M., Grigoryev, D. N., Zhang, L. Q. & Ye, S. Q. RNA-seq analysis of synovial fibroblasts brings new insights into rheumatoid arthritis. Cell Biosci. 2, 1–16 (2012).
Nota, B. et al. RNA sequencing of creatine transporter (SLC6A8) deficient fibroblasts reveals impairment of the extracellular matrix. Hum. Mutat. 35, 1128–1135 (2014).
Lahiry, P. et al. Transcriptional profiling of endocrine cerebro-osteodysplasia using microarray and next-generation sequencing. PLoS ONE 6, 1–8 (2011).
Schaum, N. et al. Single-cell transcriptomics of 20 mouse organs creates a Tabula Muris. Nature 562, 367–372 (2018).
Green, C. D. et al. A Comprehensive Roadmap of Murine Spermatogenesis Defined by Single-Cell RNA-Seq. Developmental Cell 46, 651–667 (2018).
Wei, K. et al. Notch signalling drives synovial fibroblast identity and arthritis pathology. Nature 582, 259–264 (2020).
Bartoschek, M. et al. Spatially and functionally distinct subclasses of breast cancer-associated fibroblasts revealed by single cell RNA sequencing. Nat. Commun. 9, 5150 (2018).
Wang, G. et al. Specific fibroblast subpopulations and neuronal structures provide local sources of Vegfc-processing components during zebrafish lymphangiogenesis. Nat. Commun. 11, 2724 (2020).
Muhl, L. et al. Single-cell analysis uncovers fibroblast heterogeneity and criteria for fibroblast and mural cell identification and discrimination. Nat. Commun. 11, 3953–3953 (2020).
Buechler, M. B. et al. Cross-tissue organization of the fibroblast lineage. Nature 593, 575–579 (2021).
Vorstandlechner, V. et al. Deciphering the functional heterogeneity of skin fibroblasts using single-cell RNA sequencing. FASEB J. 34, 3677–3692 (2020).
Joost, S. et al. The Molecular Anatomy of Mouse Skin during Hair Growth and Rest. Cell Stem Cell 26, 441–457.e7 (2020).
Stzepourginski, I. et al. CD34+ mesenchymal cells are a major component of the intestinal stem cells niche at homeostasis and after injury. Proc. Natl Acad. Sci. USA 114, E506–E513 (2017).
Valenta, T. et al. Wnt Ligands Secreted by Subepithelial Mesenchymal Cells Are Essential for the Survival of Intestinal Stem Cells and Gut Homeostasis. Cell Rep. 15, 911–918 (2016).
Scott, R. W., Arostegui, M., Schweitzer, R., Rossi, F. M. V. & Underhill, T. M. Hic1 Defines Quiescent Mesenchymal Progenitor Subpopulations with Distinct Functions and Fates in Skeletal Muscle Regeneration. Cell Stem Cell 25, 797–813.e9 (2019).
Elyada, E. et al. Cross-species single-cell analysis of pancreatic ductal adenocarcinoma reveals antigen-presenting cancer-associated fibroblasts. Cancer Discov. 9, 1102–1123 (2019).
Kinchen, J. et al. Structural Remodeling of the Human Colonic Mesenchyme in Inflammatory Bowel Disease. Cell 175, 372–386.e17 (2018).
Merrick, D. et al. Identification of a mesenchymal progenitor cell hierarchy in adipose tissue. Science 364, eaav2501 (2019).
Yu, Z. et al. Single-cell transcriptomic map of the human and mouse bladders. J. Am. Soc. Nephrol. 30, 2159–2176 (2019).
He, B. et al. Single-cell RNA sequencing reveals the mesangial identity and species diversity of glomerular cell transcriptomes. Nat. Commun. 12, 2141–2141 (2021).
Zeisel, A. et al. Molecular Architecture of the Mouse Nervous System. Cell 174, 999–1014 (2018).
Vanlandewijck, M. et al. A molecular atlas of cell types and zonation in the brain vasculature. Nature 554, 475–480 (2018).
Månberg, A. et al. Altered perivascular fibroblast activity precedes ALS disease onset. Nat. Med. 27, 640–646 (2021).
Liu, J. et al. A human cell type similar to murine central nervous system perivascular fibroblasts. Exp. Cell Res. 402, 112576 (2021).
Tsukui, T. et al. Collagen-producing lung cell atlas identifies multiple subsets with distinct localization and relevance to fibrosis. Nat. Commun. 11, 1920 (2020).
Melms, J. C. et al. A molecular single-cell lung atlas of lethal COVID-19. Nature 595, 114–119 (2021).
LeBleu, V. S. & Neilson, E. G. Origin and functional heterogeneity of fibroblasts. FASEB J. 34, 3519–3536 (2020).
Plikus, M. V. et al. Fibroblasts: Origins, definitions, and functions in health and disease. Cell 184, 3852–3872 (2021).
Driskell, R. R. et al. Distinct fibroblast lineages determine dermal architecture in skin development and repair. Nature 504, 277–281 (2013).
Ascensión, A. M., Fuertes-Álvarez, S., Ibañez-Solé, O., Izeta, A. & Araúzo-Bravo, M. J. Human Dermal Fibroblast Subpopulations Are Conserved across Single-Cell RNA Sequencing Studies. J. Investig. Dermatol. 141, 1735–1744.e35 (2021).
Skelly, D. A. et al. Single-Cell Transcriptional Profiling Reveals Cellular Diversity and Intercommunication in the Mouse Heart. Cell Rep. 22, 600–610 (2018).
Litviňuková, M. et al. Cells of the adult human heart. Nature 588, 466–472 (2020).
Rock, J. R. et al. Multiple stromal populations contribute to pulmonary fibrosis without evidence for epithelial to mesenchymal transition. Proc. Natl Acad. Sci. USA 108, E1475–E1483 (2011).
Peng, T. et al. Coordination of heart and lung co-development by a multipotent cardiopulmonary progenitor. Nature 500, 589–592 (2013).
Croft, A. P. et al. Distinct fibroblast subsets drive inflammation and damage in arthritis. Nature 570, 246–251 (2019).
Leigh, N. D. et al. Transcriptomic landscape of the blastema niche in regenerating adult axolotl limbs at single-cell resolution. Nat. Commun. 9, 5153 (2018).
Gerber, T. et al. Single-cell analysis uncovers convergence of cell identities during axolotl limb regeneration. Science 362, eaaq0681 (2018).
Halpern, K. B. et al. Paired-cell sequencing enables spatial gene expression mapping of liver endothelial cells. Nat. Biotechnol. 36, 962–970 (2018).
Halpern, K. B. et al. Single-cell spatial reconstruction reveals global division of labour in the mammalian liver. Nature 542, 352–356 (2017).
Deutsch, A., Feng, D., Pessin, J. E. & Shinoda, K. The impact of single-cell genomics on adipose tissue research. Int. J. Mol. Sci. 21, 1–13 (2020).
Gupta, R. K. et al. Zfp423 Expression Identifies Committed Preadipocytes and Localizes to Adipose Endothelial and Perivascular Cells. Cell Metab. 15, 230–239 (2012).
Ferguson, G. B. et al. Mapping molecular landmarks of human skeletal ontogeny and pluripotent stem cell-derived articular chondrocytes. Nat. Commun. 9, 3634 (2018).
Reynard, L. N. & Barter, M. J. Osteoarthritis year in review 2019: genetics, genomics and epigenetics. Osteoarthr. Cartil. 28, 275–284 (2020).
Vazquez-Liebanas, E. et al. Adult-induced genetic ablation distinguishes PDGFB roles in blood-brain barrier maintenance and development. J. Cereb. Blood Flow. Metab. 42, 264–279 (2021).
Armulik, A., Genové, G. & Betsholtz, C. Pericytes: developmental, physiological, and pathological perspectives, problems, and promises. Dev. Cell 21, 193–215 (2011).
Lynch, M. D. & Watt, F. M. Fibroblast heterogeneity: implications for human disease. J. Clin. Investig. 128, 26–35 (2018).
Ando, K. et al. Peri-arterial specification of vascular mural cells from naïve mesenchyme requires Notch signaling. Development 146, dev165589 (2019).
Cipleu, C. D., Palant, C. E., Sanders, K. M. & Dick, G. M. Separation of two Cl– Currents in Cultured Human and Murine Mesangial Cells: Biophysical and Pharmacological Characteristics of ICl.vol and ICl.Ca. J. Vasc. Res. 39, 426–436 (2002).
Schlondorff, D. The glomerular mesangial cell: an expanding role for a specialized pericyte. FASEB J. 1, 272–281 (1987).
Ferland-McCollough, D., Slater, S., Richard, J., Reni, C. & Mangialardi, G. Pericytes, an overlooked player in vascular pathobiology. Pharmacol. Therapeutics 171, 30–42 (2017).
Leveen, P. et al. Mice deficient for PDGF B show renal, cardiovascular, and hematological abnormalities. Genes Dev. 8, 1875–1887 (1994).
Lindahl, P. et al. Paracrine PDGF-B/PDGF-Rβ signaling controls mesangial cell development in kidney glomeruli. Development 125, 3313–3322 (1998).
Soriano, P. Abnormal kidney development and hematological disorders in PDGF b-receptor mutant mice. Genes Dev. 8, 1888–1896 (1994).
Kida, Y. & Duffield, J. S. Pivotal role of pericytes in kidney fibrosis. Clin. Exp. Pharmacol. Physiol. 38, 467–473 (2011).
Lu, Y., Ye, Y., Yang, Q. & Shi, S. Single-cell RNA-sequence analysis of mouse glomerular mesangial cells uncovers mesangial cell essential genes. Kidney Int. 92, 504–513 (2017).
Menon, R. et al. Single-cell analysis of progenitor cell dynamics and lineage specification in the human fetal kidney. Development 145, dev 164038 (2018).
Karaiskos, N. et al. A single-cell transcriptome atlas of the mouse glomerulus. J. Am. Soc. Nephrol. 29, 2060–2068 (2018).
Kupffer, K. Uber Sternzellen der Leber. Briefliche Mitteilung an Professor Waldeyer. Arch. Mikr Anat. 12, 353–358 (1876).
Pinzani, M. Hepatic stellate (ITO) cells: expanding roles for a liver-specific pericyte. J. Hepatol. 22, 700–706 (1995).
Geerts, A. History, heterogeneity, developmental biology, and functions of quiescent hepatic stellate cells. Semin. Liver Dis. 21, 311–335 (2001).
Mederacke, I. et al. Fate tracing reveals hepatic stellate cells as dominant contributors to liver fibrosis independent of its aetiology. Nat. Commun. 4, 2823 (2013).
Iwakiri, Y., Shah, V. & Rockey, D. C. Vascular pathobiology in chronic liver disease and cirrhosis—Current status and future directions. J. Hepatol. 61, 912–924 (2014).
Hellstrom, M., Kalen, M., Lindahl, P., Abramsson, A. & Betsholtz, C. Role of PDGF-B and PDGFR-beta in recruitment of vascular smooth muscle cells and pericytes during embryonic blood vessel formation in the mouse. Development 126, 3047–3055 (1999).
Dobie, R. et al. Single-Cell Transcriptomics Uncovers Zonation of Function in the Mesenchyme during Liver Fibrosis. Cell Rep. 29, 1832–1847 (2019).
Göritz, C. et al. A Pericyte Origin of Spinal Cord Scar Tissue. Science 333, 238–243 (2011).
Dias, D. O. et al. Reducing Pericyte-Derived Scarring Promotes Recovery after Spinal Cord Injury. Cell 173, 153–165 (2018).
Soderblom, C. et al. Perivascular Fibroblasts Form the Fibrotic Scar after Contusive Spinal Cord Injury. J. Neurosci. 33, 13882–13887 (2013).
Dorrier, C. E. et al. CNS fibroblasts form a fibrotic scar in response to immune cell infiltration. Nat. Neurosci. 24, 234–244 (2021).
Park, J. et al. Single-cell transcriptomics of the mouse kidney reveals potential cellular targets of kidney disease. Science 360, 758–763 (2018).
Papalexi, E. & Satija, R. Single-cell RNA sequencing to explore immune cell heterogeneity. Nat. Rev. Immunol. 18, 35–45 (2018).
Kuppe, C. et al. Decoding myofibroblast origins in human kidney fibrosis. Nature 589, 281–286 (2021).
Xie, T. et al. Single-Cell Deconvolution of Fibroblast Heterogeneity in Mouse Pulmonary Fibrosis. Cell Rep. 22, 3625–3640 (2018).
Kalluri, R. & Weinberg, R. A. The basics of epithelial-mesenchymal transition. J. Clin. Investig. 119, 1420–1428 (2009).
Zhang, W. et al. Spatial-temporal targeting of lung-specific mesenchyme by a Tbx4 enhancer. BMC Biol. 11, 111 (2013).
Tavassoli, M. & Crosby, W. H. Transplantation of marrow to extramedullary sites. Science 161, 54–56 (1968).
Friedenstein, A. J., Chailakhjan, R. K. & Lalykina, K. S. The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells. Cell Tissue Kinet. 3, 393–403 (1970).
Frenette, P. S., Pinho, S., Lucas, D. & Scheiermann, C. Mesenchymal stem cell: Keystone of the hematopoietic stem cell niche and a stepping-stone for regenerative medicine. Annu. Rev. Immunol. 31, 285–316 (2013).
Bianco, P. et al. The meaning, the sense and the significance: Translating the science of mesenchymal stem cells into medicine. Nat. Med. 19, 35–42 (2013).
Dominici, M. et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8, 315–317 (2006).
Crisan, M. et al. A Perivascular Origin for Mesenchymal Stem Cells in Multiple Human Organs. Cell Stem Cell 3, 301–313 (2008).
Thomas, H. M., Cowin, A. J. & Mills, S. J. The importance of pericytes in healing: Wounds and other pathologies. Int. J. Mol. Sci. 18, 1129 (2017).
Corselli, M., Chen, C. W., Crisan, M., Lazzari, L. & Péault, B. Perivascular ancestors of adult multipotent stem cells. Arteriosclerosis, Thrombosis, Vasc. Biol. 30, 1104–1109 (2010).
Denu, R. A. et al. Fibroblasts and Mesenchymal Stromal/Stem Cells Are Phenotypically Indistinguishable. Acta Haematologica 136, 85–97 (2016).
Driskell, R. R. & Watt, F. M. Understanding fibroblast heterogeneity in the skin. Trends Cell Biol. 25, 92–99 (2015).
Rinkevich, Y. et al. Identification and isolation of a dermal lineage with intrinsic fibrogenic potential. Science 348, aaa2151 (2015).
Abbasi, S. et al. Distinct Regulatory Programs Control the Latent Regenerative Potential of Dermal Fibroblasts during Wound Healing. Cell Stem Cell 27, 396–412.e6 (2020).
Soliman, H. et al. Pathogenic Potential of Hic1-Expressing Cardiac Stromal Progenitors. Cell Stem Cell 26, 205–220.e8 (2020).
Joe, A. W. B. et al. Muscle injury activates resident fibro/adipogenic progenitors that facilitate myogenesis. Nat. Cell Biol. 12, 153–163 (2010).
Uezumi, A., Fukada, S. I., Yamamoto, N., Takeda, S. & Tsuchida, K. Mesenchymal progenitors distinct from satellite cells contribute to ectopic fat cell formation in skeletal muscle. Nat. Cell Biol. 12, 143–152 (2010).
Pillai, I. C. L. et al. Cardiac Fibroblasts Adopt Osteogenic Fates and Can Be Targeted to Attenuate Pathological Heart Calcification. Cell Stem Cell 20, 218–232.e5 (2017).
di Carlo, S. E. & Peduto, L. The perivascular origin of pathological fibroblasts. J. Clin. Investig. 128, 54–63 (2018).
el Agha, E. et al. Mesenchymal Stem Cells in Fibrotic Disease. Cell Stem Cell 21, 166–177 (2017).
Kramann, R., Wongboonsin, J., Chang-Panesso, M., Machado, F. G. & Humphreys, B. D. Gli1+ pericyte loss induces capillary rarefaction and proximal tubular injury. J. Am. Soc. Nephrol. 28, 776–784 (2017).
Kramann, R. et al. Perivascular Gli1+ progenitors are key contributors to injury-induced organ fibrosis. Cell Stem Cell 16, 51–66 (2015).
Schneider, R. K. et al. Gli1+ Mesenchymal Stromal Cells Are a Key Driver of Bone Marrow Fibrosis and an Important Cellular Therapeutic Target. Cell Stem Cell 20, 785–800.e8 (2017).
Álvarez-Aznar, A. et al. Tamoxifen-independent recombination of reporter genes limits lineage tracing and mosaic analysis using CreERT2 lines. Transgen. Res. 29, 53–68 (2020).
Adams, T. S. et al. Single-cell RNA-seq reveals ectopic and aberrant lung-resident cell populations in idiopathic pulmonary fibrosis. Sci. Adv. 6, eaba1983 (2020).
McLellan, M. A. et al. High-Resolution Transcriptomic Profiling of the Heart during Chronic Stress Reveals Cellular Drivers of Cardiac Fibrosis and Hypertrophy. Circulation 142, 1448–1463 (2020).
Sahai, E. et al. A framework for advancing our understanding of cancer- associated fibroblasts. Nat. Rev. Cancer 20, 174–186 (2020).
Paulsson, J. & Micke, P. Prognostic relevance of cancer-associated fibroblasts in human cancer. Semin. Cancer Biol. 25, 61–68 (2014).
Karnoub, A. E. et al. Mesenchymal stem cells within tumour stroma promote breast cancer metastasis. Nature 449, 557–563 (2007).
Raz, Y. et al. Bone marrow-derived fibroblasts are a functionally distinct stromal cell population in breast cancer. J. Exp. Med. 215, 3075–3093 (2018).
Zhang, Y. et al. White adipose tissue cells are recruited by experimental tumors and promote cancer progression in mouse models. Cancer Res. 69, 5259–5266 (2009).
Dirat, B. et al. Cancer-associated adipocytes exhibit an activated phenotype and contribute to breast cancer invasion. Cancer Res. 71, 2455–2465 (2011).
Nieman, K. M. et al. Adipocytes promote ovarian cancer metastasis and provide energy for rapid tumor growth. Nat. Med. 17, 1498–1503 (2011).
Lodato, M. A. et al. Somatic mutation in single human neurons tracks developmental and transcriptional history. Science 350, 94–98 (2015).
Leung, M. L. et al. Single-cell DNA sequencing reveals a late-dissemination model in metastatic colorectal cancer. Genome Res. 27, 1287–1299 (2017).
Kester, L. & van Oudenaarden, A. Single-Cell Transcriptomics Meets Lineage Tracing. Cell Stem Cell 1–14 https://doi.org/10.1016/j.stem.2018.04.014 (2018).
VanHorn, S. & Morris, S. A. Next-Generation Lineage Tracing and Fate Mapping to Interrogate Development. Dev. Cell 56, 7–21 (2020).
Ziegenhain, C. et al. Comparative Analysis of Single-Cell RNA Sequencing Methods. Mol. Cell 65, 631–643 (2017).
Hagemann-Jensen, M. et al. Single-cell RNA counting at allele and isoform resolution using Smart-seq3. Nat. Biotechnol. 38, 708–714 (2020).
Koenitzer, J. R., Wu, H., Atkinson, J. J., Brody, S. L. & Humphreys, B. D. Single-nucleus RNA-sequencing profiling of mouse lung reduced dissociation bias and improved rare cell-type detection compared with single-cell RNA sequencing. Am. J. Respir. Cell Mol. Biol. 63, 739–747 (2020).
Hagberg, C. E. et al. Flow Cytometry of Mouse and Human Adipocytes for the Analysis of Browning and Cellular Heterogeneity. Cell Rep. 24, 2746–2756.e5 (2018).
Österreicher, C. H. et al. Fibroblast-specific protein 1 identifies an inflammatory subpopulation of macrophages in the liver. Proc. Natl Acad. Sci. 108, 308–313 (2011).
Delorey, T. M. et al. COVID-19 tissue atlases reveal SARS-CoV-2 pathology and cellular targets. Nature 595, 107–113 (2021).
He, S. et al. Single-cell transcriptome profiling an adult human cell atlas of 15 major organs. Genome Biol. 21, 294 (2020).
Acosta, J. R. et al. Single cell transcriptomics suggest that human adipocyte progenitor cells constitute a homogeneous cell population. Stem Cell Res. Ther. 8, 250 (2017).
Burl, R. B. et al. Deconstructing Adipogenesis Induced by β3-Adrenergic Receptor Activation with Single-Cell Expression Profiling. Cell Metab. 28, 300–309.e4 (2018).
Schwalie, P. C. et al. A stromal cell population that inhibits adipogenesis in mammalian fat depots. Nature 559, 103–108 (2018).
Vijay, J. et al. Single-cell analysis of human adipose tissue identifies depot- and disease-specific cell types. Nat. Metab. 2, 97–109 (2020).
Zhang, Z. et al. Dermal adipose tissue has high plasticity and undergoes reversible dedifferentiation in mice. J. Clin. Investig. 129, 5327–5342 (2019).
Chen, Z. et al. Single-cell RNA sequencing highlights the role of inflammatory cancer-associated fibroblasts in bladder urothelial carcinoma. Nat. Commun. 11, 5077 (2020).
Gao, Y. et al. CD63+ Cancer‐Associated Fibroblasts Confer Tamoxifen Resistance to Breast Cancer Cells through Exosomal miR‐22. Adv. Sci. 7, 2002518 (2020).
Wu, S. Z. et al. Stromal cell diversity associated with immune evasion in human triple‐negative breast cancer. EMBO J. 39, e104063 (2020).
Dani, N. et al. A cellular and spatial map of the choroid plexus across brain ventricles and ages. Cell 184, 3056–3074.e21 (2021).
He, L. et al. Data Descriptor: Single-cell RNA sequencing of mouse brain and lung vascular and vessel-associated cell types. Sci. Data 5, 1180160 (2018).
Shi, J. et al. Spatio-temporal landscape of mouse epididymal cells and specific mitochondria-rich segments defined by large-scale single-cell RNA-seq. Cell Discov. 7, 34 (2021).
de Bakker, D. E. M. et al. Prrx1b restricts fibrosis and promotes Nrg1-dependent cardiomyocyte proliferation during zebrafish heart regeneration. Development 148, dev198937 (2021).
Farbehi, N. et al. Single-cell expression profiling reveals dynamic flux of cardiac stromal, vascular and immune cells in health and injury. eLife 8, e43882 (2019).
DeLaughter, D. M. et al. Single-Cell Resolution of Temporal Gene Expression during Heart Development. Dev. Cell 39, 480–490 (2016).
Garvin, A. M. et al. Transient ACE (Angiotensin-Converting Enzyme) Inhibition Suppresses Future Fibrogenic Capacity and Heterogeneity of Cardiac Fibroblast Subpopulations. Hypertension 77, 904–918 (2021).
Gladka, M. M. et al. Single-Cell Sequencing of the Healthy and Diseased Heart Reveals Cytoskeleton-Associated Protein 4 as a New Modulator of Fibroblasts Activation. Circulation 138, 166–180 (2018).
Hulin, A. et al. Maturation of heart valve cell populations during postnatal remodeling. Development 146, dev173047 (2019).
Liang, D. et al. Cellular and molecular landscape of mammalian sinoatrial node revealed by single-cell RNA sequencing. Nat. Commun. 12, 287 (2021).
Liu, Z. et al. Single-cell transcriptomics reconstructs fate conversion from fibroblast to cardiomyocyte. Nature 551, 100–104 (2017).
Wang, L. et al. Single-cell dual-omics reveals the transcriptomic and epigenomic diversity of cardiac non-myocytes. Cardiovasc. Res. 118, 1548–1563 (2021).
Bahar Halpern, K. et al. Lgr5+ telocytes are a signaling source at the intestinal villus tip. Nat. Commun. 11, 1936 (2020).
Elmentaite, R. et al. Single-Cell Sequencing of Developing Human Gut Reveals Transcriptional Links to Childhood Crohn’s Disease. Dev. Cell 55, 771–783.e5 (2020).
Fawkner-Corbett, D. et al. Spatiotemporal analysis of human intestinal development at single-cell resolution. Cell 184, 810–826.e23 (2021).
Holloway, E. M. et al. Mapping Development of the Human Intestinal Niche at Single-Cell Resolution. Cell Stem Cell 28, 568–580.e4 (2021).
Hong, S. P. et al. Distinct fibroblast subsets regulate lacteal integrity through YAP/TAZ-induced VEGF-C in intestinal villi. Nat. Commun. 11, 4102 (2020).
Nanus, D. E. et al. Synovial Tissue from Sites of Joint Pain in Knee Osteoarthritis Patients Exhibits a Differential Phenotype with Distinct Fibroblast Subsets. eBioMedicine 72, 103618 (2021).
Kramann, R. et al. Parabiosis and single-cell RNA sequencing reveal a limited contribution of monocytes to myofibroblasts in kidney fibrosis. JCI Insight 3, e99561 (2018).
Lake, B. B. et al. A single-nucleus RNA-sequencing pipeline to decipher the molecular anatomy and pathophysiology of human kidneys. Nat. Commun. 10, 2832 (2019).
Affo, S. et al. Promotion of cholangiocarcinoma growth by diverse cancer-associated fibroblast subpopulations. Cancer Cell 39, 866–882 (2021).
Bhattacharjee, S. et al. Tumor restriction by type I collagen opposes tumor-promoting effects of cancer-associated fibroblasts. J. Clin. Investig. 131, e146987 (2021).
Krenkel, O., Hundertmark, J., Ritz, T., Weiskirchen, R. & Tacke, F. Single Cell RNA Sequencing Identifies Subsets of Hepatic Stellate Cells and Myofibroblasts in Liver Fibrosis. Cells 8, 503 (2019).
Massalha, H. et al. A single cell atlas of the human liver tumor microenvironment. Mol. Syst. Biol. 16, e9682 (2020).
Habermann, A. C. et al. Single-cell RNA sequencing reveals profibrotic roles of distinct epithelial and mesenchymal lineages in pulmonary fibrosis. Sci. Adv. 6, eaba1972 (2020).
Lambrechts, D. et al. Phenotype molding of stromal cells in the lung tumor microenvironment. Nat. Med. 24, 1277–1289 (2018).
Lee, J. H. et al. Anatomically and Functionally Distinct Lung Mesenchymal Populations Marked by Lgr5 and Lgr6. Cell 170, 1149–1163.e12 (2017).
Peyser, R. et al. Defining the activated fibroblast population in lung fibrosis using single-cell sequencing. Am. J. Respir. Cell Mol. Biol. 61, 74–85 (2019).
Redente, E. F. et al. Loss of Fas signaling in fibroblasts impairs homeostatic fibrosis resolution and promotes persistent pulmonary fibrosis. JCI Insight 6, e141618 (2021).
Travaglini, K. J. et al. A molecular cell atlas of the human lung from single-cell RNA sequencing. Nature 587, 619–625 (2020).
Valenzi, E. et al. Single-cell analysis reveals fibroblast heterogeneity and myofibroblasts in systemic sclerosis-associated interstitial lung disease. Ann. Rheum. Dis. 78, 1379–1387 (2019).
Wang, C. et al. Expansion of hedgehog disrupts mesenchymal identity and induces emphysema phenotype. J. Clin. Investig. 128, 4343–4358 (2018).
Yun, J. H. et al. Hedgehog interacting protein-expressing lung fibroblasts suppress lymphocytic inflammation in mice. JCI Insight 6, e144575 (2021).
Zepp, J. A. et al. Distinct Mesenchymal Lineages and Niches Promote Epithelial Self-Renewal and Myofibrogenesis in the Lung. Cell 170, 1134–1148.e10 (2017).
Rodda, L. B. et al. Single-Cell RNA Sequencing of Lymph Node Stromal Cells Reveals Niche-Associated Heterogeneity. Immunity 48, 1014–1028.e6 (2018).
Dominguez, C. X. et al. Single-Cell RNA Sequencing Reveals Stromal Evolution into LRRC15 + Myofibroblasts as a Determinant of Patient Response to Cancer Immunotherapy. Cancer Discov. 10, 232–253 (2020).
Lee, J. J. et al. Elucidation of Tumor-Stromal Heterogeneity and the Ligand-Receptor Interactome by Single-Cell Transcriptomics in Real-world Pancreatic Cancer Biopsies. Clin. Cancer Res. 27, 5912–5921 (2021).
Ligorio, M. et al. Stromal Microenvironment Shapes the Intratumoral Architecture of Pancreatic Cancer. Cell 178, 160–175.e27 (2019).
Lin, W. et al. Single-cell transcriptome analysis of tumor and stromal compartments of pancreatic ductal adenocarcinoma primary tumors and metastatic lesions. Genome Med. 12, 80 (2020).
Foster, D. S. et al. Elucidating the fundamental fibrotic processes driving abdominal adhesion formation. Nat. Commun. 11, 4061 (2020).
Joseph, D. B. et al. Single-cell analysis of mouse and human prostate reveals novel fibroblasts with specialized distribution and microenvironment interactions. J. Pathol. 255, 141–154 (2021).
de Micheli, A. J. et al. Single-cell transcriptomic analysis identifies extensive heterogeneity in the cellular composition of mouse Achilles tendons. Am. J. Physiol. Cell Physiol. 319, C885–C894 (2020).
de Micheli, A. J. et al. Single-Cell Analysis of the Muscle Stem Cell Hierarchy Identifies Heterotypic Communication Signals Involved in Skeletal Muscle Regeneration. Cell Rep. 30, 3583–3595.e5 (2020).
Giordani, L. et al. High-Dimensional Single-Cell Cartography Reveals Novel Skeletal Muscle-Resident Cell Populations. Mol. Cell 74, 609–621.e6 (2019).
Harvey, T., Flamenco, S. & Fan, C. M. A Tppp3 + Pdgfra + tendon stem cell population contributes to regeneration and reveals a shared role for PDGF signalling in regeneration and fibrosis. Nat. Cell Biol. 21, 1490–1503 (2019).
Johnson, G. L., Masias, E. J. & Lehoczky, J. A. Cellular Heterogeneity and Lineage Restriction during Mouse Digit Tip Regeneration at Single-Cell Resolution. Dev. Cell 52, 525–540.e5 (2020).
Julien, A. et al. Direct contribution of skeletal muscle mesenchymal progenitors to bone repair. Nat. Commun. 12, 2860 (2021).
Meng, S. et al. Reservoir of fibroblasts promotes recovery from limb ischemia. Circulation 142, 1647–1662 (2020).
Murach, K. A. et al. Early satellite cell communication creates a permissive environment for long-term muscle growth. iScience 24, 102372 (2021).
Xu, Z. et al. Single-cell RNA sequencing and lipidomics reveal cell and lipid dynamics of fat infiltration in skeletal muscle. J. Cachexia, Sarcopenia Muscle 12, 109–129 (2021).
Deng, C. C. et al. Single-cell RNA-seq reveals fibroblast heterogeneity and increased mesenchymal fibroblasts in human fibrotic skin diseases. Nat. Commun. 12, 3709 (2021).
Dobie, R. et al. Deciphering mesenchymal drivers of human Dupuytren’s disease at single-cell level. J. Investig. Dermatol. 142, 114–123 (2021).
Guerrero-Juarez, C. F. et al. Single-cell analysis reveals fibroblast heterogeneity and myeloid-derived adipocyte progenitors in murine skin wounds. Nat. Commun. 10, 650 (2019).
He, H. et al. Single-cell transcriptome analysis of human skin identifies novel fibroblast subpopulation and enrichment of immune subsets in atopic dermatitis. J. Allergy Clin. Immunol. 145, 1615–1628 (2020).
Layton, T. B. et al. Cellular census of human fibrosis defines functionally distinct stromal cell types and states. Nat. Commun. 11, 2768 (2020).
Leavitt, T. et al. Prrx1 Fibroblasts Represent a Pro-fibrotic Lineage in the Mouse Ventral Dermis. Cell Rep. 33, 108356 (2020).
Mahmoudi, S. et al. Heterogeneity in old fibroblasts is linked to variability in reprogramming and wound healing. Nature 574, 553–558 (2019).
Philippeos, C. et al. Spatial and Single-Cell Transcriptional Profiling Identifies Functionally Distinct Human Dermal Fibroblast Subpopulations. J. Investig. Dermatol. 138, 811–825 (2018).
Solé-Boldo, L. et al. Single-cell transcriptomes of the human skin reveal age-related loss of fibroblast priming. Commun. Biol. 3, 188 (2020).
Tabib, T., Morse, C., Wang, T., Chen, W. & Lafyatis, R. SFRP2/DPP4 and FMO1/LSP1 Define Major Fibroblast Populations in Human Skin. J. Investig. Dermatol. 138, 802–810 (2018).
Tabib, T. et al. Myofibroblast transcriptome indicates SFRP2hi fibroblast progenitors in systemic sclerosis skin. Nat. Commun. 12, 4384 (2021).
Zou, Z. et al. A Single-Cell Transcriptomic Atlas of Human Skin Aging. Dev. Cell 56, P383–P397 (2021).
Schiebinger, G. et al. Optimal-Transport Analysis of Single-Cell Gene Expression Identifies Developmental Trajectories in Reprogramming. Cell 176, 928–943.e22 (2019).
Acknowledgements
The authors are indebted to Professor Donald McDonald, Department of Anatomy, UCSF, for valuable suggestions and constructive critique on the manuscript. Work in the authors’ laboratories is supported by the Swedish Research Council, the Swedish Cancer Society, the Swedish Brain Foundation, and the Erling Persson Family Foundation (U.L., C.B.), the Knut and Alice Wallenberg Foundation (C.B.) and by Magnus Bergvalls Stiftelse (L.M.).
Author information
Authors and Affiliations
Contributions
U.L., L.M. and C.B. conceived and wrote the review. L.M. made the Figure and Box drawings.
Corresponding author
Ethics declarations
Competing interests
U.L. holds a research grant from Merck KGaA but no personal remuneration. C.B and L.M. have no competing interests.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary information
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
About this article
Cite this article
Lendahl, U., Muhl, L. & Betsholtz, C. Identification, discrimination and heterogeneity of fibroblasts. Nat Commun 13, 3409 (2022). https://doi.org/10.1038/s41467-022-30633-9
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41467-022-30633-9
- Springer Nature Limited
This article is cited by
-
Single Cell Atlas: a single-cell multi-omics human cell encyclopedia
Genome Biology (2024)
-
Distinct origin and region-dependent contribution of stromal fibroblasts to fibrosis following traumatic injury in mice
Nature Neuroscience (2024)
-
Regulation of metaplasia and dysplasia in the stomach by the stromal microenvironment
Experimental & Molecular Medicine (2024)
-
Heterogeneous fibroblasts contribute to fibrotic scar formation after spinal cord injury in mice and monkeys
Nature Communications (2024)
-
Pericytes contribute to pulmonary vascular remodeling via HIF2α signaling
EMBO Reports (2024)