摘要
目的研究大鼠骨髓间充质干细胞(BMSCs)离体分离和培养方法,探讨碱性成纤维细胞生长因子(bFGF)、表皮生长因子(EGF)、全反式维甲酸(RA)、骨形态发生蛋白-7(BMP-7)在体外诱导BMSCs向神经干细胞(NSCs)分化的作用。方法采用出生4周SD大鼠的全骨髓细胞进行培养,传至第3代时,分为3组:A组,bFGF+EGF+RA进行诱导分化;B组,BMP-7进行诱导分化;C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神经元特异性烯醇化酶(NSE)单抗鉴定。结果A组诱导7 d后有部分细胞具备神经元样细胞形态,胞体呈锥形或圆形,有较长单极或多极的突起,有NSE阳性细胞表达。而B组、C组细胞仍为典型的成纤维细胞形态,无NSE阳性细胞。结论bFGF、EGF和RA可以联合诱导BMSCs向NSCs分化,并可调控神经元分化,而BMP-7在此种诱导条件下未见明显的诱导BMSCs向NSCs分化的作用。
Objective To investigate the isolating and culturing methods of rat bone marrow stromal cells （BMSCs）in vitro and to study whether adult BMSCs could be induced to differentiate into neural stem cells （NSCs） and neurons by basic fibroblast growth factor（ bFGF）, epidermal growth factor（ EGF）, all-transretinoic acid（RA） and bone morphogenetic protein-7 （BMP-7 ）. Methods Adult marrow stromal cells were cultured from bone marrow of four week-old SD rats. bFGF ＋ EGF ＋ RA （ group A） and BMP-7 （ group B） were used to induce the third-generation BMSCs, and contrasted with the cells that had not been induced. Phase contrast microscope was used to observe the differentiation of BMSCs daily. Immunohistochemistry technique was used to identify the differentiated BMSCs with neuron specific enolase （NSE）. Results After seven days of culture, some BMSCs in grupe A appeared typical morphological neuronal traits. Cell bodies appeared refractile, conelike or spherical with long processes, NSE was expressed in group A. The cells in group B and group C still had no NSE expression. Conclusion Combination of bFGF, EGF and RA can induce BMSCs to differentiate into NSCs and neurons, and can regulate the differentiation and promote the maturation of neurons. But BMP-7 can not induce BMSCs to differentiate into NSCs and neurons at this condition.
出处
《现代医学》
2008年第2期102-105,共4页
Modern Medical Journal
