Comparing The Biologic Characters Of Adipose Tissue-derived Mesenchymal Stem Cells Of Different Persons

Objective Transplantation of adult mesenchymal stem cells has been proposed as a hopeful strategy for cardiac repair following myocardial damage. Adipose tissue-derived mesenchymal stem cells (ADMSCs) as a novel seed cell for cytological engineering have been gradually accepted. Now there are two main mechanisms to explain the functions of stem cells in treating myocardial infarction: differentiation and paracrine. Scientists have found that cell sex is a variable that considerably influences the regeneration abilities of muscle-derived stem cells'(MDSCs) and found that the female MDSCs regenerated skeletal muscle more efficiently in mice. In this study we try to found whether the cell sex or cell age will influence the human ADMSCs' biologic characters, including proliferation, differentiation, paracrine and anti-apoptosis abilities.Methods According to the aims mentioned above, this study was divided into two parts.Part I Cells were obtained by digesting adipose tissue of human beings with trypsin and Collagenase, and then cultured in vitro. The third passage cells were determined with immunocytochemistry methods for surface molecules CD13,CD44,CD45,HLA-DR,VWF, and induced with lineage-specific induction factors to fat, bone and cardiac muscle which were determined with different staining methods of Oil red O, alkaline Phosphatase and FCM method.Part II We used the MTT method to compare the growth curves of the third passage of different persons' ADMSCs. And the ELISA method was used for comparing the abilities of secretion of HGF and VEGF of the stem cells. The differentiation and anti-apoptosis abilities of the h-ADMSCs of different persons were evaluated through the FCM method.ResultsWe had successfully obtained spindle-shaped cells from human's adipose tissue. Immunocytochemistry showed that most of cells were positive for CD13,CD44 and negative for CD45,HLA-DR,VWF. They could differentiate into adipogenic, osteogenic and cardiac muscle cells in the presence of lineage-specific induction factors.The MTT experiment showed the OD values of the female ADMSCs(F-ADMSCs) were significantly higher than the male's in the 5, 7, 9 days. Induced by the 10 umol/L 5-aza for 24 hours, F-ADMSCs showed a higher expression of TNT than M-ADMSCs (27.3%±2.4% VS 18.9%±2.5%, P<0.01) by FCM method. And the younger ADMSCs were easier to be induced. The ELISA result showed the F-ADMSCs secreted more VEGF(1582±132 pg per 10⁶cells VS 1265±133 pg per 10⁶cells, P<0.01) and HGF(13316±1404 pg per 10⁶cells VS 11790±1541 pg/10⁶个细胞P<0.01) than M-ADMSCs. After induced by 500umol/L H₂O₂ for 30 minutes, F-ADMSCs presented less apoptosis rate than M-ADMSCs (18.68%±1.35% VS 25.21%±1.57%, P<0.01). And the younger ADMSCs were easier to been induced to apoptosis.Conclusion1. Large quantity of ADMSCs with multilineage differentiation potential can be easily obtained from human's adipose tissue by enzyme digestion.2. There are sex differences in the proliferation, differentiation, paracrine and anti-apoptosis abilities of the h-ADMSCs. And age also plays an important part in the h-ADMSCs' differentiation and anti-apoptosis abilities.