摘要
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× 10 5/ 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。
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;2.5×10 5 cells/cm 2 was the best planting density;the best first medium changing was the fifth day at primary culture, DMEM medium was better than α-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.
出处
《中国修复重建外科杂志》
CAS
CSCD
2004年第3期220-224,共5页
Chinese Journal of Reparative and Reconstructive Surgery
基金
国家自然科学基金资助项目 (30 1 71 0 0 7)~~
