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• W2415067981 abstract "It appears that much progress has been made since Alexander Friedenstein's first descriptions of bone marrow (BM) stromal cells, variably described in the following years as mesenchymal stem or stromal cells (MSCs) and featuring high in vitro replicative capacity as well as osteogenic and chondrogenic differentiation potential.1-4 However, despite decades of research leading to a fair understanding of role of MSCs in maintenance of the hematopoietic niche, it is the obvious heterogeneity of MSCs that has been posing an extraordinary challenge for our understanding of MSCs biology and their translation to the clinic. MSCs can be isolated from basically all organs and tissues of the adult organism as well as from umbilical cord blood and tissue, Wharton's jelly, placenta, and amniotic fluid or membrane.6, 7 Although sharing certain properties such as fibroblastoid morphology, adherent growth, and suppression of immune cell proliferation in vitro,8, 9 differences can be detected regarding differentiation potential, marker expression or miRNA profiling between MSCs derived from the BM, adipose tissue (AT), or cord blood.8, 10 It remains to be clarified whether these differences relate to the functional context of the tissue origin of native MSCs and their actual function in situ, for example, stromal support within the BM niche and metabolic or immune homeostasis within the AT (see below). Analyzing MSC markers in situ confirm the presence of distinct MSC subpopulations in different microanatomic sites in the human BM.11 Specifically, reticular stromal cells, composed of CD10+CD73+CD146+ adipose stromal cells and CD10+CD31+CD146+GD2+ fibroblastoid reticular cells, can be distinguished from CD10+CD73+CD140b+CD271+ round stromal cells, CD73+CD140b+CD271+GD2+ bone lining cells, and perivascular cells.11 Moreover, variable in vivo differentiation capacity of transplanted BM-MSCs point toward conservation of MSC heterogeneity after transplantation.12 This suggests different subtypes and probably different function of MSCs in situ in relation to their respective niche. In addition to the most commonly used MSC isolation method, that is, adherence to tissue culture–treated plastic surfaces, other matrix-based isolation technologies have been applied such as collagen I/III or fibrin matrices.13 Additionally, an increasing repertoire of selection technologies, such as positive selection by FACS or MACS using specific antibodies against CD49a, CD271, TNAP, or MSC-specific aptamers as well as negative selection of hematopoietic cells offer options to enrich and to subculture more defined MSC phenotypes.13, 14 Specific culture conditions, for example low oxygen atmosphere, feeder layer, or growth factor cocktails in the medium can select for MSC entities such as multipotent adult progenitor cells, BM-isolated adult multilineage inducible cells, or very small embryonic-like stem cells.15 Moreover, it is certainly possible that culture in bioreactors or 3D culture systems may affect MSC biology compared to “standard” culture technology, that is, 2D tissue culture–treated plastic flasks.16 When it became clear that data from MSC studies vary, a “minimal standard” for MSC characterization was implemented, that is, plastic adherence, trilineage (adipogenic, osteogenic, chondrogenic) in vitro differentiation and a minimalistic panel of markers assessed by flow cytometry (CD73+, CD90+, CD105+, CD34–, CD45–). This first attempt of standardization was a remarkable improvement to compare MSC data obtained from different laboratories and therefore, a significant step forward in the field. However, reports that mesenchymal progenitor cells could be detected even in the nonadherent fraction of the BM,17 that markers other than the above mentioned “minimal panel” might define MSC subpopulations with function and from other tissues than BM,18, 19 and that donor age and sex can influence MSC function20 highlight the need for further MSC characterization and a better understanding of their heterogeneity. It is also striking that the level of heterogeneity changes within different passages, suggesting differential marker expression in single cells or dynamics in the composition of subpopulations.21 To address these issues, the following strategies have been developed to tackle MSC heterogeneity (Table 1): Phenotyping over time of multiple clones Live cell imaging In vitro screening studies Sort and enrich subpopulations Establish optimal culture conditions for subpopulations Study subpopulation biology in vivo/in situ Dissect mechanisms (loss/gain of function studies) Develop in vitro assays that predict in vivo efficacy and safety Identify candidate endpoints based on mechanistic studies (e.g., marker or expression profile a,b,c corresponding to function x) Correlate with clinical data Siegel and colleagues20 analyzed human BM-MSCs from more than 50 donors that were isolated and cultured under standardized conditions. Statistical analyses of these cells revealed positive correlations of marker expression (CD10, CD71, CD106, CD119, CD146, CD166, CD271) to clonogenic potential of MSCs, whereas other markers (CD140b, Galectin1) correlated negatively with secretion of hepatocyte growth factor. Interestingly, MSCs obtained from female donors showed greater immunomodulatory and clonogenic potential, whereas donor sex did not affect differentiation capacity of MSCs in vitro. As outlined above, MSC subpopulations can be sorted, subcultured, and characterized by a variety of descriptive and functional assays. In particular, analysis of CD271+ sorted MSCs revealed interesting functional insights into MSC subpopulation biology. Specifically, CD271+ BM-MSCs feature greater osteogenic differentiation potential and an increased capacity to suppress T-cell proliferation in vitro compared to nonsorted BM-MSCs.22, 23 Moreover, CD31–CD34+CD45– sorted dermal mesenchymal progenitor cells show distinct differentiation potential compared to other subpopulations in the dermis.24 The fact that sorted CD271+ BM-MSC clones feature different immunomodulatory properties in vitro23 highlights the need for more comprehensive characterization, ideally at the single-cell level. More recent studies have applied screening technologies for multiparametric assessment of MSCs. Walmsley et al.25 analyzed AT-MSCs by a flow cytometry screening panel comprised of 242 antibodies before and after adipogenic or osteogenic differentiation. With this approach, markers for osteogenic differentiation and adipogenic differentiation could be identified. Microfluidic single-cell gene expression analysis is an intriguing tool to address MSCs heterogeneity. This technology allows the quantitative assessment of expression of dozens of genes in hundreds of individual cells thereby achieving statistically robust gene expression clustering of subpopulations within cell preparations.26 Applying this technology, Duscher and coworkers27 identified distinct clusters (subpopulations) in AT-MSC preparations from young and aged mice featuring functional differences such as support of vasculogenesis and wound healing. In summary, advanced characterization of MSCs, and especially MSC subpopulations, is mandatory to tackle MSC heterogeneity, to advance our understanding of MSC biology and to sustainably develop MSC therapies and meaningful potency assays. The authors have disclosed no conflicts of interest." @default.
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• W2415067981 date "2016-04-01" @default.
• W2415067981 modified "2023-10-14" @default.
• W2415067981 title "Characterization of mesenchymal stem or stromal cells: tissue sources, heterogeneity, and function" @default.
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