论文部分内容阅读
目的观察大鼠骨髓基质细胞(rBMSCs)的生长特点及诱导条件下分化成神经细胞的能力,并对其机制进行初步探讨。方法以密度梯度离心分离骨髓基质细胞,在神经干细胞培养液中培养,采用四唑盐(MTT)法观察在培养液中添加碱性成纤维细胞生长因子(bFGF)、表皮生长因子(EGF)对BMSCs增殖的影响;观察添加脑源性神经生长因子(BDNF)、神经生长因子(NGF)和维甲酸(RA)对rBMSCs的诱导分化情况;采用免疫组织化学法(ABC)检测诱导后的细胞表达神经元特异性烯醇化酶(NSE)、神经元核蛋白(NeuN)和胶质原性纤维酸性蛋白抗体(GFAP)等特异性标志物的情况;以流式细胞分选确定神经元的比例。结果bFGF和EGF能在体外促进rBMSCs增殖,BDNF、NGF和RA能诱导rBMSCs来源的神经干细胞(NSCs)表达NSE、GFAP等特异性标志物。结论EGF、bFGF、BDNF、NGF、RA及适宜的培养液可使rBMSCs定向转化为NSCs,获得足够的目的细胞,进而分化为神经元样和神经胶质样细胞。
OBJECTIVE: To observe the growth characteristics of rat bone marrow stromal cells (rBMSCs) and their ability to differentiate into neurons under induction conditions, and to explore the mechanism of them. Methods Bone marrow stromal cells were isolated by density gradient centrifugation and cultured in neural stem cell culture medium. The effects of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) BMSCs proliferation was observed. The induced differentiation of rBMSCs was observed by adding BDNF, NGF and RA. The induced cells were detected by immunohistochemistry (ABC) Neuron-specific enolase (NSE), neuronal nucleoprotein (NeuN) and glial fibrillary acidic protein antibody (GFAP) and other specific markers; the proportion of neurons determined by flow cytometry. Results bFGF and EGF could promote the proliferation of rBMSCs in vitro. BDNF, NGF and RA could induce NSCs derived from rBMSCs to express specific markers such as NSE and GFAP. CONCLUSIONS: EGF, bFGF, BDNF, NGF, RA and appropriate culture medium can transform rBMSCs into NSCs and obtain enough target cells to differentiate into neuron-like and glial-like cells.