Promotion of human adipose-derived stem cell proliferation mediated by exogenous nucleosides.
Adult stem cells are becoming the best option for regenerative medicine because they have low tumourigenic potential and permit autologous transplantation, even without in vitro culture. Our objectives were to evaluate the effects of exogenous nucleosides on the proliferation of hASCs (human adipose-derived stem cells), with or without co-treatment with 5-aza (5-azacytidine), and to analyse the expression of lamin A/C during cardiomyocyte differentiation of these cells. We isolated hASCs from human lipoaspirates that were positive for mesenchymal stem cell markers. We found that 5-aza induces a dose-dependent inhibition of hASC proliferation [IC50 (inhibitory concentration 50): 5.37 microM], whereas exogenous nucleosides significantly promote the proliferation of hASCs and partially revert the antiproliferative effect of the drug. Multipotentiality of isolated hASCs was confirmed by adipogenic, osteogenic and cardiomyogenic induction. 5-Aza-induced cells expressed cardiac troponins I and T and myosin light chain 2, myocardial markers that were directly correlated with lamin A/C expression. Our results support the importance of the nucleoside supplementation of media to improve conditions for the expansion and maintenance of hASCs in culture. In addition, the quantification of lamin A/C expression appears to be a good marker for the characterization of cardiomyocyte differentiation of stem cells that has rarely been used.
Department of Human Anatomy and Embryology, University of Granada, 18012 Granada, Spain.
Fernando
F
Rodríguez-Serrano
Pablo
P
Alvarez
Octavio
O
Caba
Manuel
M
Picón
Juan A
JA
Marchal
Macarena
M
Perán
José
J
Prados
Consolación
C
Melguizo
Ana R
AR
Rama
Houria
H
Boulaiz
Antonia
A
Aránega
D000273:Adipose Tissue / Q000166:cytology*; D001374:Azacitidine / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D034904:Lamin Type A / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D009705:Nucleosides / Q000494:pharmacology*; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism
D015415:Biomarkers; D034904:Lamin Type A; D009705:Nucleosides; D001374:Azacitidine