小鼠孤雌胚胎干细胞直接向神经细胞诱导分化的实验研究

Direct differentiation of mouse parthenogenetic stem cells into neural cells without embryonic body culture in vitro

目的探讨小鼠孤雌胚胎干细胞在体外不经过拟胚体(EB)阶段,直接诱导分化为神经细胞的可行性。方法采用阶段诱导的方法,首先由孤雌胚胎干细胞向神经前体细胞定向分化,通过巢蛋白(nestin)免疫荧光细胞化学染色对神经前体细胞进行鉴定。在此基础上,撤除丝裂原,加入5%胎牛血清,观察已分化的神经前体细胞能否进一步分化为神经细胞,并对分化出的细胞进行免疫荧光细胞化学鉴定。结果经选择培养基培养3 d后的孤雌干细胞绝大多数可诱导为神经前体细胞,nestin呈阳性。加入血清进一步诱导,可分化出β-Ⅲ-tubline阳性的神经细胞。结论小鼠孤雌胚胎干细胞在体外不经EB培养阶段,可直接诱导分化为神经细胞,并且前期可得到大量的神经前体细胞。

Objective To investigate the feasibility of direct differentiation of mouse parthenogenetic stem cells into neural cells without embryonic body culture in vitro. Methods The method of phase induction was used to induce direct differentiation from parthenogenetic stem cells to neural precursors which were identified by nestin immunofluorescence staining. Sequential culture was applied for the further differentiation into neural cells with the removal of mitogen and addition of 5 % fetal bovine serum. The characteristics of the further induced cells were identified by staining. Results Large amounts of parthenogenetic stem cells were differentiated into neural precursors, which were nestin-positive in the selected medium on the 3rd day. Also β-Ⅲ-tubhne-positive neural cells could be induced from the neural precursors by further culture. Conclusion Mouse parthenogenetic stem cells could be directly differentiated into neural cells without embryonic body culture in vitro and induce large amounts of neural precursors.