Abstract
The dermis of adult human skin contains a physiologically heterogeneous population of fibroblasts that interact to produce its unique architecture and that participate in inflammatory and wound repair functions in vivo. This heterogeneity has been well documented for fibroblasts located in the superficial papillary dermis and the deep reticular dermis. However, the existence of diverse fibroblast subpopulations within a given region of the dermis has not been explored. In this study, fibroblast cultures have been established from the superficial dermis following enzymatic dissociation of the tissue. These fibroblasts have been cloned by limiting dilution and initially selected on the basis of morphology and proliferation kinetics. Fibroblasts in some of the clones selected for study express α-smooth muscle actin, a myofibroblast characteristic. Significant differences for fibroblast clones obtained from the same piece of skin have been observed with regard to their rate of collagen lattice contraction, their ability to organize a fibronectin matrix, their release of specific growth factors/cytokines into culture medium, and their response to interleukin-1α. These differences in both morphological and physiological characteristics indicate that the superficial papillary dermis contains a heterogeneous population of fibroblasts. This heterogeneity might indicate that diverse subpopulations of fibroblasts are required to interact in both homeostatic and pathological situations in skin.
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Akagi A, Tajima S, Ishibashi A, Yamoguchi N, Nagai Y (1999) Expression of type XVI collagen in human skin fibroblasts: enhanced expression in fibrotic skin diseases. J Invest Dermatol 113:246–250
Azzarone B, Macieira-Coelho A (1982) Heterogeneity of the kinetics of proliferation within human skin fibroblastic cell populations. J Cell Sci 57:177–187
Bahar MA, Bauer B, Tredget EE, Ghahary A (2004) Dermal fibroblasts from different layers of human skin are heterogeneous in expression of collagenase and type I and III procollagen mRNA. Wound Repair Regen 12:75–182
Balin AK, Fisher AJ, Anzelone M, Leong I, Allen RG (2002) Effects of establishing cell cultures and cell culture conditions on the proliferative life span of human fibroblasts isolated from different tissues and donors of different ages. Exp Cell Res 274:275–287
Bayreuther K, Rodemann HP, Francz PI, Maier K (1988a) Differentiation of fibroblast stem cells. J Cell Sci Suppl 10:115–130
Bayreuther K, Rodemann HP, Hommel R, Dittmann K, Albiez M, Francz PI (1988b) Human skin fibroblasts in vitro differentiate along a terminal cell lineage. Proc Natl Acad Sci USA 85:5112–5116
Blomme EAG, Zhou H, Kartsogiannis V, Capen CC, Rosol TJ (1999) Spatial and temporal expression of parathyroid hormone-related protein during wound healing. J Invest Dermatol 112:788–795
Botstein GR, Sherer GK, Leroy EC (1982) Fibroblast selection in scleroderma. An alternative model of fibrosis. Arthritis Rheum 25:189–195
Boxman I, Lowik C, Aarden L, Ponec M (1993) Modulation of IL-6 production and IL-1 activity by keratinocyte-fibroblast interaction. J Invest Dermatol 101:316–324
Boxman ILA, Ruwhof C, Boerman OC, Lowik CWGM, Ponec M (1996) Role of fibroblasts in the regulation of proinflammatory interleukin IL-1, IL-6, and IL-8 levels induced by keratinocyte-derived IL-1. Arch Dermatol Res 288:391–398
Chomarat P, Banchereau J, Davoust J, Palucka AK (2000) IL-6 switches the differentiation of monocytes from dendritic cells to macrophages. Nat Immunol 1:510–514
Clark RAF (1990) Fibronectin matrix deposition and fibronectin receptor expression in healing and normal skin. J Invest Dermatol 94:128s–134s
Cormack DH (1987) The integumentary system. In: Ham’s histology, 9th edn. Lippencott Company, Philadephia, pp 450–474
Coulomb B, Dubertret L (2002) Skin cell culture and wound healing. Wound Repair Regen 10:109–112
Danen EH, Yamada KM (2001) Fibronectin, integrins, and growth control. J Cell Physiol 189:1–13
Darby I, Skalli O, Gabbiani G (1990) α-Smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab Invest 63:21–29
Derdak S, Penney DP, Keng P, Felch ME, Brown D, Phipps RP (1992) Differential collagen and fibronectin production by Thy 1+ and Thy 1− lung fibroblast subpopulations. Am J Physiol 263:L282–L290
Desmoulière A, Rubbia-Brandt L, Abdiu A, Walz T, Macieira-Coelho A, Gabbiani G (1992) α-Smooth muscle actin is expressed in a subpopulation of cultured and cloned fibroblasts and is modulated by γ-interferon. Exp Cell Res 201:64–73
Desmoulière A, Chaponnier C, Gabbiani G (2005) Tissue repair, contraction, and the myofibroblast. Wound Repair Regen 13:7–12
Eckes B, Zigrino P, Kessler D, Holtkotter O, Shephard P, Mauch C, Krieg T (2000) Fibroblast-matrix interactions in wound healing and fibrosis. Matrix Biol 19:325–332
ffrench-Constant C, Van De Water L, Dvorak HF, Hynes RO (1989) Reappearance of an embryonic pattern of fibronectin splicing during wound healing in the adult rat. J Cell Biol 109:903–914
Fries KM, Blieden T, Looney RJ, Sempowski GD, Silvera MR, Willis RA, Phipps RP (1994) Evidence of fibroblast heterogeneity and the role of fibroblast subpopulations in fibrosis. Clin Immunol Immunopathol 72:283–292
Fritsch C, Orian-Rousseaul V, Lefebvre O, Simon-Assmann P, Reimund JM, Duclos B, Kedinger M (1999) Characterization of human intestinal stromal cell lines: response to cytokines and interactions with epithelial cells. Exp Cell Res 248:391–406
Gailit J, Clark RAF (1994) Wound repair in the context of extracellular matrix. Curr Opin Cell Biol 6:717–725
Gallucci RM, Sloan DK, Heck JM, Murray AR, O’Dell SJ (2004) Interleukin 6 indirectly induces keratinocyte migration. J Invest Dermatol 122:764–772
Germain L, Jean A, Auger FA, Garrel DR (1994) Human wound healing fibroblasts have greater contractile properties than dermal fibroblasts. J Surg Res 57:268–273
Goldring SR, Stephenson ML, Downie E, Krane SM, Korn JH (1990) Heterogeneity in hormone responses and patterns of collagen synthesis in cloned dermal fibroblasts. J Clin Invest 85:798–803
Grossman RM, Krueger J, Yourish D, Granelli-Piperno A, Murphy DP, May LT, Kupper TS, Sehgal, PB, Gottlieb AB (1989) Interleukin 6 is expressed in high levels in psoriatic skin and stimulates proliferation of cultured human keratinocytes. Proc Natl Acad Sci USA 86:6367–6371
Harper RA, Grove G (1979) Human skin fibroblasts derived from papillary and reticular dermis: differences in growth potential in vitro. Science 204:526–527
Jahoda CAB, Reynolds AJ, Chaponnier C, Forester JC, Gabbiani G (1991) Smooth muscle α-actin is a marker for hair follicle dermis in vivo and in vitro. J Cell Sci 99:627–636
Kessler-Becker D, Krieg T, Eckes B (2004) Expression of proinflammatory markers by human dermal fibroblasts in a three-dimensional culture model is mediated by an autocrine interleukin-1 loop. Biochem J 379:351–358
Limeback H, Sodek J, Aubin JE (1982) Variation in collagen expression by cloned periodontal ligament cells. J Periodontal Res 18:242–248
Maas-Szabowski N, Fusenig NE (1996) Interleukin-1-induced growth factor expression in postmitotic and resting fibroblasts. J Invest Dermatol 107:849–855
Maas-Szabowski N, Stark H-J, Fusenig NE (2000) Keratinocyte growth regulation in defined organotypic cultures through IL-1-induced keratinocyte growth factor expression in resting fibroblasts. J Invest Dermatol 114:1075–1084
Metz CN (2003) Fibrocytes: a unique cell population implicated in wound healing. Cell Mol Life Sci 60:1342–1350
Nakaoka H, Miyauchi S, Miki Y (1995) Proliferating activity of dermal fibroblasts in keloids and hypertrophic scars. Acta Derm Venereol 75:102–104
Nowinski D, Lysheden A-S, Gardner H, Rubin K, Gerdin B, Ivarsson M (2004) Analysis of gene expression in fibroblasts in response to keratinocyte-derived factors in vitro: potential implications for the wound healing process. J Invest Dermatol 122:216–221
Penney DP, Keng PC, Derdak S, Phipps RP (1992) Morphological and functional characteristics of subpopulations of murine lung fibroblasts grown in vitro. Anat Rec 232:432–443
Rodemann HP, Bayreuther K, Francz PI, Dittmann K, Albiez M (1989) Selective enrichment and biochemical characterization of seven human skin fibroblast cell types in vitro. Exp Cell Res 180:84–93
Schafer IA, Pandy M, Feguson R, Davis BR (1985) Comparative observation of fibroblasts derived from the papillary and reticular dermis of infants and adults: growth kinetics, packing density at confluence and surface morphology. Mech Ageing Dev 31:275–293
Schmitt-Gräff A, Desmoulière A, Gabbiani G (1994) Heterogeneity of myofibroblasts phenotypic features: an example of fibroblastic cell plasticity. Virchows Arch 425:3–24
Schönherr E, Beavan LA, Hausser H, Kresse H, Culp LA (1993) Differences in decorin expression by papillary and reticular fibroblasts in vivo and in vitro. Biochem J 290:893–899
Schwarzbauer JE, Sechler JL (1999) Fibronectin fibrillogenesis; a paradigm for extracellular matrix assembly. Curr Opin Cell Biol 11:622–627
Sechler JL, Schwarzbauer JE (1998) Control of cell cycle progression by fibronectin matrix architecture. J Biol Chem 273:25533–25536
Sempowski GD, Borrello MA, Blieden TM, Barth RK, Phipps RP (1995) Fibroblast heterogeneity in the healing wound. Wound Repair Regen 3:120–131
Smith RS, Smith TJ, Blieden TM, Phipps RP (1997) Fibroblasts as sentinel cells. Synthesis of chemokines and regulation of inflammation. Am J Pathol 151:317–322
Smola H, Thiekotter G, Fusenig NE (1993) Mutual induction of growth factor gene expression by epidermal-dermal cell interaction. J Cell Biol 122:417–429
Sorrell JM, Caplan AI (2004) Fibroblast heterogeneity: more than skin deep. J Cell Sci 117:667–675
Sorrell JM, Baber MA, Caplan AI (1996) Construction of a bi-layered dermal equivalent containing human papillary and reticular dermal fibroblasts: use of fluorescent vital dyes. Tissue Eng 2:39–49
Sorrell JM, Carrino DA, Baber MA, Asselineau D, Caplan AI (1999) A monoclonal antibody which recognizes a glycosaminoglycan epitope in both dermatan sulphate and chondroitin sulphate proteoglycans of human skin. Histochem J 31:549–559
Sorrell JM, Baber MA, Brinon L, Carrino DA, Seavolt M, Asselineau D, Caplan AI (2003) Production of a monoclonal antibody, DF-5, that identifies cells at the epithelial-mesenchymal interface in normal human skin. APN/CD13 is an epithelial-mesenchymal marker in skin. Exp Dermatol 12:315–323
Sorrell JM, Baber MA, Caplan AI (2004) Site-matched papillary and reticular human dermal fibroblasts differ in their release of specific growth factors/cytokines and in their interaction with keratinocytes. J Cell Physiol 200:134–145
Sottile J, Hocking DC, Swiatek PJ (1998) Fibronectin matrix assembly enhances adhesion-dependent cell growth. J Cell Sci 111:2933–2943
Szabowski A, Maas-Szabowski N, Andrecht S, Kolbus A, Schorpp-Kristner M, Fusenig NE, Angel P (2000) c-Jun and JunB antagonistically control cytokine-regulated mesenchymal-epidermal interaction in skin. Cell 103:745–755
Tajima S, Izumi T (1996) Differential in vitro responses of elastin expression to basic fibroblast growth factor and transforming growth factor β1 in upper middle, and lower dermal fibroblasts. Arch Dermatol Res 288:753–756
Turksen K, Kupper TS, Degenstein L, Williams I, Fuchs E (1992) Interleukin 6: insights to its function in skin by overexpression in transgenic mice. Proc Natl Acad Sci USA 89:5068–5072
Waelti ER, Inaebnit SP, Rast HP, Hunziker T, Limat A, Braathen LR, Wiesmann U (1992) Co-culture of human keratinocytes on post-mitotic human dermal fibroblast feeder cells: production of large amounts of interleukin 6. J Invest Dermatol 98:805–808
Wang H-J, Pieper J, Peters F, Blitterswijk CA van, Lamme EN (2004) Improved enzymatic isolation of fibroblasts for the creation of autologous skin substitutes. In Vitro Cell Dev Biol 40A:268–277
Werner S, Smola H (2001) Paracrine regulation of keratinocyte proliferation and differentiation. Trends Cell Biol 11:143–146
Westergren-Thorsson G, Sime P, Jordana M, Gauldie J, Särnstrand B, Malmström A (2004) Lung fibroblast clones from normal and fibrotic subjects differ in hyaluronan and decorin production and rate of proliferation. Int J Biochem Cell Biol 36:1573–1584
Wu C, Kelvens VM, O’Toole TE, McDonald JA, Ginsberg MH (1995) Integrin activation and cytoskeletal interaction are essential for the assembly of a fibronectin matrix. Cell 83:715–724
Yang L, Scott PG, Dodd C, Medina A, Jiao H, Shankowsky HA, Ghahary A, Tredget EE (2005) Identification of fibrocytes in postburn hypertrophic scar. Wound Repair Regen 13:398–404
Zucali J, Dinarello CA, Oblon D, Gross MA, Anderson L, Weiner RS (1986) Interleukin 1 stimulates fibroblasts to produce granulocyte-macrophage colony-stimulating activity and prostaglandin E2. J Clin Invest 77:1857–1863
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We thank Dr. Thomas McCormick, Department of Dermatology, Case Western Reserve University, for providing the dermatomed tissue.
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We thank L’Oréal Life Sciences for providing funding for these studies.
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Sorrell, J.M., Baber, M.A. & Caplan, A.I. Clonal characterization of fibroblasts in the superficial layer of the adult human dermis. Cell Tissue Res 327, 499–510 (2007). https://doi.org/10.1007/s00441-006-0317-y
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DOI: https://doi.org/10.1007/s00441-006-0317-y