Objective:At present, human umbilical cord mesenchymal stem cells (hucMSC) have been widely used in basic research and clinical trials in treatment of many diseases, but how to obtain a large number and high quality o...Objective:At present, human umbilical cord mesenchymal stem cells (hucMSC) have been widely used in basic research and clinical trials in treatment of many diseases, but how to obtain a large number and high quality of stable and reliable mesenchymal stem cells for clinical application is still a scientific research needs to be resolved. Since each laboratory used different culture methods, the productivity of the different cultures may vary. Therefore, the purpose of this study is to optimize a stable and reliable hucMSC culture procedure. Methods: Mesenchymal stem cells derived from human umbilical cord were isolated and cultured in different culture medium. By comparing the morphology, passage, proliferation and using flow cytometry to detect the immunophenotypes of the cells, optimized cultural method was selected.Results: By comparison of the time of cells migrating out from the tissue, cell growth status, cell number and expression of CD90 andCD73 of the hucMSCs, we found that X-VIVO15+10% fetal bovine serum group, X-VIVO15+2% fetal bovine serum and 2% UltroserG group, and MSC basal medium+UltraGRO-Advanced group had an earlier migration time from the umbilical cord and shorter confluence time to cover 80%-90% of the cuturing flask. The Huc-MSCs cultured with MSC basal medium+2% UltraGRO-Advanced group revealed the fastest proliferation. The expression of CD90 and CD73 in Huc-MSCs cultured with MSC basal medium+2% UltraGRO-Advanced group were above 90%, while the expression of CD90 and CD73 in other groups were below 90%. The expression of CD34 and CD45 surface antigens were low in huc-MSCs from all the groups.Conclusion:In summary, MSC basal medium-UltraGRO-Advanced group was the optimal culture medium for human umbilical cord mesenchymal stem cells.展开更多
基金National Natural Science Foundation(:81570157)Shandong Province Medical and Health Technology Development Project(2013ws0291).
文摘Objective:At present, human umbilical cord mesenchymal stem cells (hucMSC) have been widely used in basic research and clinical trials in treatment of many diseases, but how to obtain a large number and high quality of stable and reliable mesenchymal stem cells for clinical application is still a scientific research needs to be resolved. Since each laboratory used different culture methods, the productivity of the different cultures may vary. Therefore, the purpose of this study is to optimize a stable and reliable hucMSC culture procedure. Methods: Mesenchymal stem cells derived from human umbilical cord were isolated and cultured in different culture medium. By comparing the morphology, passage, proliferation and using flow cytometry to detect the immunophenotypes of the cells, optimized cultural method was selected.Results: By comparison of the time of cells migrating out from the tissue, cell growth status, cell number and expression of CD90 andCD73 of the hucMSCs, we found that X-VIVO15+10% fetal bovine serum group, X-VIVO15+2% fetal bovine serum and 2% UltroserG group, and MSC basal medium+UltraGRO-Advanced group had an earlier migration time from the umbilical cord and shorter confluence time to cover 80%-90% of the cuturing flask. The Huc-MSCs cultured with MSC basal medium+2% UltraGRO-Advanced group revealed the fastest proliferation. The expression of CD90 and CD73 in Huc-MSCs cultured with MSC basal medium+2% UltraGRO-Advanced group were above 90%, while the expression of CD90 and CD73 in other groups were below 90%. The expression of CD34 and CD45 surface antigens were low in huc-MSCs from all the groups.Conclusion:In summary, MSC basal medium-UltraGRO-Advanced group was the optimal culture medium for human umbilical cord mesenchymal stem cells.
