Abstract
Placenta and amniotic membrane emerge as important sources of stem cells, since it is possible to perform their study with non-invasive methods. Also, is possible to obtain higher numbers of stem cells from these tissues compared with other sources. Several studies have reported that multipotent adult stem cells represent an attractive stem cell source for regenerative medicine and transplantation therapy, since these cells have a high degree of plasticity and multi-lineage differentiation potential. Among adult stem cells, mesenchymal stem cells (MSC) arise since they are able of both supporting hematopoiesis and differentiating into mesoderm, endoderm and ectoderm cells and are known to be weakly immunogenic and to exhibit immunomodulatory properties, which is important to escape to the immunological defence mechanisms and to suppress several functions. Diverse studies associated the MSC heterogeneity with their differential potential. MSC have different characteristics related to the source of isolation, therefore they can differ, not only, in terms of phenotype, but also morphology, ultrastructure and even function. Although MSC phenotype is well described in the literature, the absence of a single marker expressed only by this cell type could be a problem for the isolation of a homogeneous stem cell population and for the identification of their therapeutic potential. In this sense, recent efforts have been made in order to isolate more homogeneous cell populations, avoiding contamination by other cellular types, and to develop a set of markers for the characterization of this stem cell type. In this context, this chapter gives an overview of protocols for isolation, a proposal for their characterization, and differentiation of mesenchymal stem cells from human amniotic membrane (hAMSC) in order to potentiate its application to clinical practice.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Parolini O, Alviano F, Bagnara GP et al (2008) Concise review – isolation and characterization of cells from human term placenta: outcome of the first international workshop on placenta derived stem cells. Stem Cells 26:300–311
Bosio A, Huppert V, Donath S et al (2009) Isolation and enrichment of stem cells. Adv Biochem Eng Biotechnol 114:23–72
Mamede AC, Carvalho MJ, Abrantes AM et al (2012) Amniotic membrane: from structure and functions to clinical applications. Cell Tissue Res 349:447–458
Dominici M, Le Blanc K, Mueller I et al (2006) Minimal criteria for defining multipotent mesenchymal stromal cells: the international society for cellular therapy position statement. Cytotherapy 8:315–317
Schrepfer S, Deuse T, Lange C et al (2007) Simplified protocol to isolate, purify and culture expand mesenchymal stem cells. Stem Cells Dev 16:105–108
Harichandan A, Bühring H-J (2011) Prospective isolation of human MSC. Best Pract Res Clin Haematol 24:25–36
Soncini M, Vertua E, Gibelli L et al (2007) Isolation and characterization of mesenchymal cells from human fetal membranes. J Tissue Eng Regen Med 1:296–305
Barbati A, Mameli MG, Sidoni A, Di Renzo GC (2012) Amniotic membrane: separation of amniotic mesoderm from amniotic epithelium and isolation of their respective mesenchymal stromal and epithelial cells. Curr Protoc Stem Cell Biol 20:1E.8.1–1E.8.15
Kim J, Kang HM, Kim H et al (2007) Ex vivo characteristics of human amniotic membrane-derived stem cells. Cloning Stem Cells 9:581–594
Alviano F, Fossati V, Marchionni C et al (2007) Term amniotic membrane is a high throughput source for multipotent mesenchymal stem cells with the ability to differentiate into endothelial cells in vitro. BMC Dev Biol 7:1–14
Koo BK, Park IY, Kim J et al (2012) Isolation and characterization of chorionic mesenchymal stromal cells from human full term placenta. J Korean Med Sci 27:857–863
Leyva-Leyva M, Barrera L, López-Camarillo C et al (2013) Characterization of mesenchymal stem cell subpopulations from human amniotic membrane with dissimilar osteoblastic potential. Stem Cells Dev 22:1275–1287
Chamberlain G, Fox J, Ashton B, Middleton J (2007) Concise review: mesenchymal stem cells – their phenotype, differentiation capacity, immunological features and potential for homing. Stem Cells 25:2739–2749
Grogan SP, Miyaki S, Asahara H et al (2009) Mesenchymal progenitor cell markers in human articular cartilage: normal distribution and changes in osteoarthritis. Arthritis Res Ther 11:R85
Sivasubramaniyan K, Lehnen D, Ghazanfari R et al (2012) Phenotypic and functional heterogeneity of human bone marrow– and amnion-derived MSC subsets. Ann N Y Acad Sci 1266:94–106
Davies D (2007) Cell sorting by flow cytometry. In: Macey M (ed) Flow Cytom. Princ. Appl. Humana Press, Totowa, pp 257–276
Ibrahim S, van den Engh G (2007) Flow cytometry and cell sorting. Adv Biochem Eng Biotechnol 106:19–39
Van Dongen J, Lhermitte L, Böttcher S et al (2012) EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukemia 26:1908–1975
Pedreira CE, Costa ES, Arroyo ME et al (2008) A multidimensional classification approach for the automated analysis of flow cytometry data. IEEE Trans Biomed Eng 55:1155–1162
Pedreira CE, Costa ES, Barrena S et al (2008) Generation of flow cytometry data files with a potentially infinite number of dimensions. Cytom A 73A:834–846
Laranjeira P, Ribeiro A, Mendes S et al (2011) Immunophenotypic characterization of normal bone marrow stem cells. In: Schmid I (ed) Flow cytom. – recent perspect. InTech, Rijeka, pp 457–478
Ibrahim SF, van den Engh G (2003) High-speed cell sorting: fundamentals and recent advances. Curr Opin Biotechnol 14:5–12
Ryan JM, Pettit AR, Guillot PV et al (2013) Unravelling the pluripotency paradox in fetal and placental mesenchymal stem cells: oct-4 expression and the case of the emperor’s new clothes. Stem Cell Rev Rep 9:408–421
Matikainen T, Laine J (2005) Placenta: an alternative source of stem cells. Toxicol Appl Pharmacol 207:544–549
Mihu CM, Ciuca DR, Soritau O et al (2009) Isolation and characterization of mesenchymal stem cells from the human amniotic membrane. Rom J Morphol Embryol 50:73–77
Allegra A, Altomare R, Curcio P et al (2013) Gene expression of stem cells at different stages of ontological human development. Eur J Obstet Gynecol Reprod Biol 170:381–386
Roubelakis MG, Trohatou O, Anagnou NP (2012) Amniotic fluid and amniotic membrane stem cells: marker discovery. Stem Cells Int 107836:1–9
Tsai M-S, Hwang S-M, Chen K-D et al (2007) Functional network analysis of the transcriptomes of mesenchymal stem cells derived from amniotic fluid, amniotic membrane, cord blood and bone marrow. Stem Cells 25:2511–2523
Baharvand H, Heidari M, Ebrahimi M et al (2007) Proteomic analysis of epithelium-denuded human amniotic membrane as a limbal stem cell nich. Mol Vis 13:1711–1721
Hopkinson A, McIntosh RS, Shanmuganathan V et al (2006) Proteomic analysis of amniotic membrane prepared for human transplantation: characterization of proteins and clinical implications. J Proteome Res 5:2226–2235
Perin L, Sedrakyan S, Da Sacco S, De Filippo R (2008) Characterization of human amniotic fluid stem cells and their pluripotential capability. Methods Cell Biol 86:85–99
Díaz-Prado S, Muiños-López E, Hermida-Gómez T et al (2011) Isolation and characterization of mesenchymal stem cells from human amniotic membrane. Tissue Eng C Methods 17:49–59
Díaz-Prado S, Muiños-López E, Hermida-Gómez T et al (2010) Multilineage differentiation potential of cells isolated from the human amniotic membrane. J Cell Biochem 111:846–857
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Lopes, A., Grãos, M., Paiva, A. (2015). Isolation and Characterization of Mesenchymal Stem Cells from Amniotic Membrane. In: Mamede, A., Botelho, M. (eds) Amniotic Membrane. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-9975-1_12
Download citation
DOI: https://doi.org/10.1007/978-94-017-9975-1_12
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-017-9974-4
Online ISBN: 978-94-017-9975-1
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)