人骨髓间质干细胞的优化培养

目的探索人骨髓间质干细胞(mesenchymal stem cells,MSCs)的优化培养条件.方法应用不同条件,研究原代培养方法、接种密度、首次换液时间、培养基、血清浓度及种类对细胞生长的影响,并以选定的条件培养、扩增MSCs,扩增后向成骨细胞和软骨细胞诱导.结果在其它条件不变的前提下,密度梯度分离法优于全骨髓法,2.5×105/cm2是人MSCs原代培养的适宜接种密度,原代第5天首次换液为最佳换液时间,DMEM培养基优于α-MEM培养基,血清C为适宜的血清,10%为适宜的血清浓度.所选条件培养的MSCs可在体外扩增15代以上,形态保持不变,并具有良好的分化潜能.结论建立了人MSCs的体外优化培养条件,为其在组织工程方面的应用作出了新的探索.

OPTIMAL CULTURE OF HUMAN BONE MARROW MESENCHYMAL STEM CELLS

OBJECTIVE: To optimize the culture conditions of the human bone marrow mesenchymal stem cells (hMSCs). METHODS: The influence of the primary culture method, planting density, the time of the first medium changing, culture medium and serum concentration on growth of the hMSCs were analyzed. RESULTS: When the other conditions were the same, the density gradient isolation was better than whole-marrow isolation; 2.5 x 10(5) cells/cm2 was the best planting density; the best first medium changing was the fifth day at primary culture, DMEM medium was better than alpha-MEM, serum C was the best of four serums compared, 10% was the suitable serum concentration. The hMSCs under the optimal conditions could expand over 15 passages, remaining their normal modality and differentiation potentials. CONCLUSION: The optimal culture condition of the hMSCs is established and it is a new investigation on application of hMSCs to tissue engineering.