Defining essential stem cell characteristics in adipose-derived stromal cells extracted from distinct anatomical sites
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The discovery of adipose-derived stromal cells (ASCs) has created many opportunities for the development of patient-specific cell-based replacement therapies. We have isolated multiple cell strains of ASCs from various anatomical sites (abdomen, arms/legs, breast, buttocks), indicating widespread distribution of ASCs throughout the body. Unfortunately, there exists a general lack of agreement in the literature as to their “stem cell” characteristics. We find that telomerase activity and expression of its catalytic subunit in ASCs are both below the levels of detection, independent of age and culturing conditions. ASCs also undergo telomere attrition and eventually senesce, while maintaining a stable karyotype without the development of spontaneous tumor-associated abnormalities. Using a set of cell surface markers that have been promoted to identify ASCs, we find that they failed to distinguish ASCs from normal fibroblasts, as both are positive for CD29, CD73 and CD105 and negative for CD14, CD31 and CD45. All of the ASC isolates are multipotent, capable of differentiating into osteocytes, chondrocytes and adipocytes, while fibroblasts show no differentiation potential. Our ASC strains also show elevated expression of genes associated with pluripotent cells, Oct-4, SOX2 and NANOG, when compared to fibroblasts and bone marrow-derived mesenchymal stem cells (BM-MSCs), although the levels were lower than induced pluripotent stem cells (iPS). Together, our data suggest that, while the cell surface profile of ASCs does not distinguish them from normal fibroblasts, their differentiation capacity and the expression of genes closely linked to pluripotency clearly define ASCs as multipotent stem cells, regardless of tissue isolation location.
KeywordsASC iPS Pluripotent Stem cell Telomere Telomerase
Special thanks to Dr. Stephen Chen and the Department of Surgery for assisting with the procurement of lipoaspirate and abdominoplasty specimens. This study was supported in part by the Department of Pathology (S.E.H. and L.W.E.), the Department of Human and Molecular Genetics (S.E.H.), K01CA105050 (L.W.E.) from the National Cancer Institute, W81XWH-09-1-0500 (L.W.E.) from the Department of Defense Breast Cancer Research Program and the Jeffress Memorial Trust (L.W.E.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Cancer Institute or the National Institutes of Health. Flow cytometry was supported in part by the Massey Cancer Center core grant, NIH Grant P30 CA16059.
All authors agree to the format and content of the manuscript and none have any commercial affiliations or associations that would contemplate a conflict of interest for the data presented.
- Abdallah BM, Haack-Sørensen M, Burns JS, Elsnab B, Jakob F, Hokland P, Kassem M (2005) Maintenance of differentiation potential of human bone marrow mesenchymal stem cells immortalized by human telomerase reverse transcriptase gene despite extensive proliferation. Biochem Biophys Res Comm 326:527–538PubMedCrossRefGoogle Scholar
- Astori G, Vignati F, Bardelli S, Tubio M, Gola M, Albertini V, Bambi F, Scali G, Castelli D, Rasini V, Soldati G, Moccetti T (2007) In vitro and multicolor phenotypic characterization of cell subpopulations identified in fresh human adipose tissue stromal vascular fraction and in the derived mesenchymal stem cells. J Transl Med 5:55PubMedCrossRefGoogle Scholar
- Barch M (1991) The AGT cytogenetics manual, 3 rdth edn. Lippencott Williams and Wilkins, Philadelphia, pp 263–265Google Scholar
- Rebelatto CK, Aguiar AM, Moretão MP, Senegaglia AC, Hansen P, Barchiki F, Oliveira J, Martins J, Kuligovski C, Mansur F, Christofis A, Amaral VF, Brofman PS, Goldenberg S, Nakao LS, Correa A (2008) Dissimilar differentiation of mesenchymal stem cells from bone marrow, umbilical cord blood, and adipose tissue. Exp Biol Med (Maywood) 233:901–913CrossRefGoogle Scholar
- Rooney D, Czepulkowski B (1992) Human cytogenetics: A practical approach, vol II malignancy and acquired abnormalities, 2nd edn. Oxford University Press, New York, pp 198–200Google Scholar
- Torsvik A, Røsland GV, Svendsen A, Molven A, Immervoll H, McCormack E, Lønning PE, Primon M, Sobala E, Tonn JC, Goldbrunner R, Schichor C, Mysliwietz J, Lah TT, Motaln H, Knappskog S, Bjerkvig R (2010) Spontaneous malignant transformation of human mesenchymal stem cells reflects cross-contamination: putting the research field on track. Cancer Res 70:6393–6396PubMedCrossRefGoogle Scholar