不同血清浓度下人骨髓间充质干细胞定向分化为神经元样细胞的实验研究  

The research of inducing hBMSCs to differentiate to neural like cells in different density serum

作　　者：玉石[1] 刘卫平[1] 刘阳[1] 龙乾发[1] 王孝安[1] 陈晓燕[1] 李娟[1] 

机构地区：[1]第四军医大学附属西京医院神经外科,西安710032

出　　处：《立体定向和功能性神经外科杂志》2009年第5期274-278,共5页Chinese Journal of Stereotactic and Functional Neurosurgery

摘　　要：目的探讨不同血清浓度对体外诱导人骨髓间充质干细胞向神经样细胞分化的影响。方法采集成人志愿者的骨髓细胞进行体外培养,传至第3代时,分为6组:第一组(无血清诱导组),10ng/mlbFGF+10ng/mlEGF预诱导72h后,0.1umol/LRA、10ng/mlGDNF+10ng/mlBDNF正式诱导,其余各组分别加入1%FBS、2%FBS、5%FBS、10%FBS、20%FBS。在倒置显微镜下每6～8h观察、记录BMSCs的诱导分化情况,并应用荧光免疫组化技术对分化后的细胞进行鉴定。结果各组经正式诱导1d后有部分BMSC开始收缩,加血清组的BM-SC生长速度快,部分细胞出现神经元及神经胶质细胞形态。7d完成分化后,2%血清组NSE显色最多,GFAP最少,而20%血清组NSE显色最少,2%血清组细胞平均分化率为26.5%,20%血清组平均分化率为12.5%。结论体外诱导BMSCs向神经元样细胞分化时,2%血清浓度可起促进作用,而高浓度血清则起抑制作用。Objective To investigate the effect of different FBS densitytoward inducing human bone marrow stromal cells（BMSCs）differentiate into neural like cells in vitro.Methods Adult marrow stromal cells were cultured from bone marrow of the volunteer.After 10ng/ml bFGF ＋ EGF induced for 72h,0.1umol/LRA ＋10ng/ml BDNF＋10ng/ml GDNF without FBS（group1）were used to induce the third generation BMSCs,and the other groups were added fetal bull blood serum with densityof 1%、2%、5%、10% and 20% separately.Phase contrast microscope was used to observe the differentiation of BMSCs every 68h.and fluorescence immunohistochemistry technique was used to identify the differentiated BMSCs.Results After 1 day of culture,some BMSCs appeared to contract,the cells in FBS groups grows faster than none FBS group,part of these cells turned into typical morphological neuronal and glia traits.7days after differentiation completed,2%FBS group cells were discover expressed NSE more than other groups after fluorescence immunohistochemistry test and expressed GFAP least.The average differentiate rate of 2%FBS group was 26.5%,and the 20%FBS group was 12.5%.Conclusion 2%FBS can promote BMSCs differentiate into neuron-like cells in vitro,while the high densityFBS may inhibit the differentiation.

关 键 词：骨髓间充质干细胞 胎牛血清 神经元 

分 类 号：Q343[生物学—遗传学]