Surface protein characterization of human adipose tissue‐derived stromal cells

Abstract

Human bone marrow stromal cells are a multipotent population of cells capable of differentiating into a number of mesodermal lineages as well as supporting hematopoeisis. Their distinct protein and gene expression phenotype is well characterized in the literature. Human adipose tissue presents an alternative source of multipotent stromal cells. In this study, we have defined the phenotype of the human adipose tissue-derived stromal cells in both the differentiated and undifferentiated states. Flow cytometry and immunohistochemistry show that human adipose tissue-derived stromal cells have a protein expression phenotype that is similar to that of human bone marrow stromal cells. Expressed proteins include CD9, CD10, CD13, CD29, CD34, CD44, CD 49(d), CD 49(e), CD54, CD55, CD59, CD105, CD106, CD146, and CD166. Expression of some of these proteins was further confirmed by PCR and immunoblot detection. Unlike human bone marrow-derived stromal cells, we did not detect the STRO-1 antigen on human adipose tissue-derived stromal cells. Cells cultured under adipogenic conditions uniquely expressed C/EBPalpha and PPARdelta, two transcriptional regulators of adipogenesis. Cells cultured under osteogenic conditions were more likely to be in the proliferative phases of the cell cycle based on flow cytometric analysis of PCNA and Ki67. The similarities between the phenotypes of human adipose tissue-derived and human bone marrow-derived stromal cells could have broad implications for human tissue engineering.

2010 Nicolas Andry Award: Multipotent Adult Stem Cells from Adipose Tissue for Musculoskeletal Tissue Engineering

“…This population of cells can be further expanded and purified on tissue culture plastic, yielding approximately 10 9 cells after 2 weeks of expansion. During expansion, our studies show that ASCs do not exhibit markers of hematopoietic stem cells (eg, CD45 and CD14) but display a similar marker surface profile to marrow derived MSCs [32,83], although, it generally is recognized there are no specific markers readily available to identify nonhematopoietic stem cells. Nevertheless, ASCs express the stromal markers CD9, CD10, CD29, CD44, CD73, CD90, and CD166, and with increasing passage, the expression of these markers increases while the presence of hematopoietic markers declines [57,61].…”

In vitro cardiomyogenic differentiation of adipose‐derived stromal cells using transforming growth factor‐β1

“…These results are similar to previous studies. 21,[27][28][29] ADSCs treated with TGF-b1 showed cardiac-specific marker expression that was absent from untreated ADSCs. Western blot analysis showed expression of cardiac-specific marker expression, troponin I, in ADSCs cultured with TGFb1, but not in ADSCs cultured without TGF-b1 (Figure 2).…”

“…15,21 ADSCs express multiple CD marker antigens, such as CD29, CD44, CD90, and CD105, similar to other mesenchymal stem cells (MSCs). 21,28 ADSC multipotency has been demonstrated in their ability to differentiate into various lineages including adipocytes, osteoblasts, chondrocytes, and endothelial cells. 15,27,[31][32][33] Contracting cells with CMC features were seen during adipose stromal cell culture due to spontaneous differentiation, but the frequency was very low (0.02-0.07%).…”

Phenotypic Characterization and In Vivo Localization of Human Adipose-Derived Mesenchymal Stem Cells

“…Therefore, the results of flow cytometric evaluation or cell sorting could vary greatly among different specimens, persons, or passages. For instance, the expression of well-known surface markers including CD105, CD73, and CD90 can change according to the cell passage number (Gronthos et al, 2001;Lin et al, 2010;McIntosh et al, 2006;Mitchell et al, 2006) In order to resolve some of this confusion, some groups have attempted in vivo identification using immunofluorescence markers including ones specific for PDGFR, α-SMA, CD146, and NG2. Despite these efforts, precise identification of hADMSCs is still not possible (Crisan et al, 2008;Lin et al, 2008).…”