诱导成鼠骨髓源神经干细胞的实验研究

Study of induction of neural stem cells derived from adult rat bone marrow

目的:探讨体外诱导成鼠骨髓源神经干细胞的可行性。方法:以含20ng/ml碱性成纤维生长因子(basicfibroblastgrowthfactor,bFGF)与表皮生长因子(epidermalgrowthfactor,EGF)的无血清培养液培养成鼠骨髓细胞,待细胞球形成后进行传代培养与单细胞克隆试验,以含100ng/mlbFGF培养液诱导分化,细胞免疫组化染色进行细胞鉴定。结果:骨髓细胞在诱导条件下形成表达Nestin抗原的细胞球,亦表达造血干细胞标志性抗原CD90。分离成单细胞后很快形成新的细胞球,并可分化出神经元与胶质细胞。结论:骨髓细胞可在体外诱导为能自我增殖的神经干细胞。

Objective:To investigate induction of neural stem cells from adult rat bone marrow in vitro.Method:Bone marrow cells were cultivated in serum-free medium with 20ng/ml basic fibroblast growth factor(bFGF) and epidermal growth factor(EGF).After cellular spheres forming,primary spheres were mechanically dissociated and were cultivated or performed single-cell cloning test.Spheres were differentiated in medium with 100ng/ml bFGF,then immunocytochemistry test were performed.Result:In inductive situation,bone marrow could give rise to cellular spheres.The spheres expressed nestin,a neural stem cell marker,as well as CD90,a marker of hematopoietic stem cells.New spheres could soon form after being dissociated,and spheres could differationate into neurons and glial cells.Conclusion:Bone marrow cells can be induced to neural stem cells with self-proliferation characteristics.