Culture and identification of rhesus monkey bone marrow mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Studies on rhesus monkey bone marrow mesenchymal stem cells (BMSCs) are limited,and the differentiation potential of juvenile rhesus monkey BMSCs remains unknown.OBJECTIVE:To investigate the biological characteristics of rhesus monkey BMSCs isolated and cultured in vitro,and to identify the growth curve and phenotype of BMSCs.METHODS:BMSCs from juvenile rhesus monkeys were isolated and cultured by density gradient centrifugation and the whole bone marrow adherence methods.BMSCs at passages 2 and 7 were taken to draw cell growth curves by MTS method.Flow cytometry was used to detect apoptosis of passage 3 BMSCs and to detect expression of cell surface markers,CD29,CD34,CD45,CD90,CD147,in passage 4 BMSCs.Osteogenic and adipogenic abilities of passage 3 BMSCs were also detected.RESULTS AND CONCLUSION:The BMSCs at passages 2 and 7 were incubated at first 2 days,logarithmically grew at 3-7 days,and then proliferated slowly at 8 days.The BMSCs at passage 2 showed stronger proliferation than those at passage 7.At passage 3,BMSCs were negative for CD34 and CD45,but positive for CD90,CD29 and CD147.Results from the fiow cytometry showed that there were 73.83％ living cells,21.76％ dead cells,and 4.27％ apoptotic cells.BMSCs could differentiate into osteoblasts and adipocytes under osteogenic and adipogenic induction,respectively.To conclude,BMSCs from the rhesus monkey have strong proliferation and multilineage differentiation abilities in vitro.