Cell Surface and Transcriptional Characterization of Human Adipose‐Derived Adherent Stromal (hADAS) Cells

Abstract

Adult human subcutaneous adipose tissue contains cells with intriguing multilineage developmental plasticity, much like marrow-derived mesenchymal stem cells. Putative stem or progenitor cells from fat have been given many different names in the literature, reflecting an early and evolving consensus regarding their phenotypic characterization. The study reported here used microarrays to evaluate over 170 genes relating to angiogenesis and extracellular matrix in undifferentiated, early-passage human adipose-derived adherent stromal (hADAS) cells isolated from three separate donors. The hADAS populations unanimously transcribed 66% of the screened genes, and 83% were transcribed by at least two of the three populations. The most highly transcribed genes relate to functional groupings such as cell adhesion, matrix proteins, growth factors and receptors, and proteases. The transcriptome of hADAS cells demonstrated by this work reveals many similarities to published profiles of bone marrow mesenchymal stem cells (MSCs).In addition, flow analysis of over 24 hADAS cell surface proteins (n = 7 donors) both confirms and expands on the existing literature and reveals strong intergroup correlation, despite an inconsistent nomenclature and the lack of standardized protocols for cell isolation and culture. Finally, based on flow analysis and reverse transcription polymerase chain reaction studies, our results suggest that hADAS cells do not express several proteins that are implicated as markers of "stemness" in other stem cell populations, including telomerase, CD133, and the membrane transporter ABCG2. Stem Cells 2005;23:412-423

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“…Both cell types express CD105, CD73, CD90 and lack the haematopoietic lineage markers c-kit, CD14, CD11b, CD34, CD45, CD79, CD19 and HLA-DR. These markers are considered the minimal prerequisite to describe MSC (Katz et al, 2005;Schaffler and Buchler, 2007). In 2006, MSC were defined by the International Society for Cellular Therapy as a plastic-adherent cell population with the following surface marker profile: CD13 + , CD44 + , CD90 + , CD73 + and CD105 + , CD14 − , CD11b − , CD79 − , CD34 − , CD45 − and HLA-DR − (Dominici et al, 2006).…”

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“…Also, haematopoietic cells were derived using mouse adipose tissue-derived stroma vascular fraction (Charriere et al, 2003;Cousin et al, 2003). Studies were undertaken to characterize ADSCs; these reports showed that the phenotype and genotype at a transcriptional level are quite similar to those of BMSCs (DeUgarte et al, 2003;Katz et al, 2005). However, differentiation potentials might be different.…”

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“…ASCs were isolated as previously described [17,18]. Briefly, samples were washed, enzymatically dissociated, and filtered to remove debris [18].…”