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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 serum concentrations on the differentiation of human bone marrow mesenchymal stem cells into neuron-like cells in vitro. Methods Adult bone marrow cells from adult volunteers were cultured in vitro and divided into 6 groups at the third passage: group Ⅰ (serum-free group), pre-induction with 10ng / ml bFGF + 10ng / ml EGF for 72h, 0.1umol / LRA , 10ng / mlGDNF + 10ng / mlBDNF formal induction, and the remaining groups were added 1% FBS, 2% FBS, 5% FBS, 10% FBS, 20% FBS. Under inverted microscope, the differentiation and differentiation of BMSCs were observed every 6 ~ 8h, and the differentiated cells were identified by fluorescence immunohistochemistry. Results After being formally induced for 1 day, some BMSCs began to contract, and BM-SC in serum-added group grew faster. Some neurons and glial cells appeared in some cells. After 7 days of differentiation, NSE developed the most in 2% serum group with the least in GFAP, while the staining of NSE in 20% serum group was the least. The average differentiation rate of NSE in 2% serum group was 26.5% and that in 20% serum group was 12.5%. Conclusions 2% serum concentration can promote the differentiation of BMSCs to neuron-like cells in vitro, while high concentration serum can inhibit the proliferation of BMSCs.