bFGF与EGF诱导BMSCs向神经干细胞分化的观察

目的 探讨碱性成纤维细胞生长因子（bFGF）、表皮生长因子（EGF）在体外条件下对骨髓基质干细胞（BMSCs）向神经千细胞（NSCs）和神经样细胞分化的诱导作用。方法 采用出生4周左右大鼠的全骨髓细胞进行培养，传至第3代时，分为两组：实验组细胞加入bFGF和EGF，进行诱导分化；对照组不加因子继续培养。在倒置显微镜下每日观察，记录BMSCs的诱导分化情况，并应用免疫组化技术对细胞进行兔抗巢蛋白（Nestin）单抗、兔抗神经元特异性烯醇化酶（NSE）单抗、兔抗胶质纤维酸性蛋白（GFAP）单抗鉴定。结果 实验组诱导7d后有部分细胞具备神经元样细胞形态，胞体锥形或圆形，有较长单极或多极的突起，均有Nestin、NSE、GFAP阳性细胞表达，且3种细胞数量差别有显著性（t=3．266～6．099，P〈0．05）。而对照组细胞仍为典型的成纤维细胞形态，无Nestin、NSE、GFAP阳性细胞。结论 bFGF、EGF可以诱导BMSCs向NSCs分化，并可调控神经元分化，促进神经元细胞成熟。

BMSCs DIFFERENTIATION INTO NSCs INDUCED BY bFGF AND EGF

Objective To study whether adult bone marrow stromal cells(BMSCs) could be induced to differentiate into neural stem cells (NSCs) and neurons by basic fibroblast growth factor(bFGF) and epidermal growth factor(EGF). Methods(Adult marrow stromal) cells were cultured from bone marrow of four-week-old Wistar rats.bFGF and EGF were used to induce the thirdgeneration BMSCs,and contrasted with the cells that had not been induced.A phase contrast microscope was used to observe the differentiation of BMSCs,daily.Immunohistochemistry technique was used to identify the differentiated BMSCs with Nestin,neuron specific enolase(NSE) and glial fibrillary acid protein(GFAP). Results After seven days of culture,some BMSCs appeared typical morphological neuronal traits: the refractile and cone-like or spherical cell bodies and long processes,expressed Nestin or NSE or GFAP.The three kinds of cells showed significant difference,but the cells in the control group still had no Nestin or NSE or GFAP expression. Conclusion bFGF and EGF could induce BMSCs to differentiate into NSCs and neurons,and might regulate the differentiation and promote the maturation of neurons.