体外诱导大鼠骨髓间质细胞转化为神经细胞

Transdifferentiation of adult rat bone marrow stromal cells to neural cells in vitro

目的体外培养骨髓间质细胞,探索大鼠骨髓间质细胞体外诱导及向神经细胞方向分化。方法分离大鼠骨髓间质细胞培养、扩增,检查细胞活力。应用脑神经营养因子(BDNF)、β-巯基乙醇(-βME)对细胞诱导分化,行大体观察,6 h后免疫组织化鉴定神经丝蛋白(NF-200)、神经烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP),并用RT-PCR鉴定NF-200、NSE和GFAP的mRNA表达情况。结果用BDNF、-βME单独或者共同诱导3 h,骨髓间质细胞形态类似神经细胞,6 h后神经细胞标记蛋白和mRNA表达阳性,而且BDNF+-βME组诱导率最高。结论BDNF和-βME能协同刺激大鼠骨髓间质细胞转化并分化为神经细胞。

Objective To observe the growth and differentiation of cultured rat bone marrow cells （MSCs） to neural cells. Methods Purified MSCs were induced by β-mercaptoethanol（β-ME）alone or combination of brain derived neurotrophic factor（BDNF） with β-ME. After 6 h incubation, neuron special endolase （NSE）, neurofilament （NF-200） and glial fibrillary acidic protein （GFAP） were detected by immunohistochemostry,and mRNAs of NSE, NF-200 and GFAP were detected with reverse transcriptase-polymerase chain reaction （RT-PCR） and the cells with vitality were observed. Results After induction with β-ME or BDNF alone or together, some MSCs exhibited the phenotypes of neural cells. After 6 h induction, NSE and NF-200 positive cells increased significantly. It was the same in the mRNA. Furthermore, the addition of BDNF and β-ME caused the maximal variation. Conclusion Combination of β-ME with BDNF may synergistically promote the transformation of MSCs and differentiation into neurons.