同种异体血清在体外培养骨髓间充质干细胞的作用研究

目的]适当的血清浓度可维持骨髓间充质干细胞(MSCs)的体外培养,血清浓度过低不利于细胞的生长,而高浓度血清则易引起细胞分化。而血清的不同来源对MSCs的培养同样产生影响。传统的培养方法用胎牛血清作为营养支持,临床应用时存在潜在的风险。本研究探讨同种异体血清在体外培养人骨髓间充质干细胞(hBM-SCs)的可行性。方法]无菌条件下收集人骨髓悬液,分离hBMSCs,分别用含胎牛血清和人血清的培养基培养。分别测定2种方法培养的hBMSCs细胞生长曲线;采用流式细胞仪检测分析培养的hBMSCs表面抗原类型;并将培养的hBMSCs诱导分化为软骨细胞及神经细胞,诱导后的软骨细胞及神经细胞采用免疫组化染色分析。结果]2种方法培养的间充质干细胞表面抗原CD29、CD44、CD105阳性,CD34、CD45、CD106以及HLA-DR阴性,表达强度无显著性差异;人血清培养的hBMSCs细胞生长速度快于胎牛血清培养组,但分化效率低于后者。结论]通过生长特性、表面抗原表达以及分化潜能等方面的对比研究,人血清培养hBMSCs与胎牛血清培养差别不大,可以作为一种适合临床的安全培养方法。

Experiment study of the role of allogeneic serum in the culture of human bone marrow mesenchymal stem cells in vitro

Objective]To investigate if allogeneic human serum(alloHS)could replace fetal bovine serum(FBS)for the expansion of human bone marrow mesenchymal stem cells(hBMSCs)in vitro.Method]Human BM-MSCs were isolated either from spongy bone residues obtained during hip surgery or from washed filters used during bone marrow graft processing for allogeneic bone marrow transplantation.We culture human bone marrow mesenchymal stem cells were cultivated using alloHS and FBS,then observe its growth character,antigen presentation and differentiation potency through condition medium directional induction were observed.Result]Isolated MSCs were positive for the markers CD105,CD29 and CD44 and negative for typical hematopoietic and endothelial markers CD34,CD45,CD106 and HLA-DR.The choice of serum affected hMSCs at several different levels.Firstly,hMSCs in alloHS proliferated markedly faster than hMSCs in FBS.Secondly,hMSCs in FBS differentiated more rapidly toward mesenchymal lineages compared with hMSCs in alloHS.Conclusion]hMSCs may be expanded rapidly in alloHS in the absence of growth factors,this may be a safely culture method suitable for clinical demand.