维生素A酸、锌等诱导大鼠骨髓间质干细胞分化为神经元样细胞的研究

in vitro Differentiation of Rat Bone Mesenchymal Stem Cells into Neuron-like Cells Induced by VitaminA Acid etc.

目的 探讨维生素 A酸、锌及大鼠损伤脊髓提取液联合诱导大鼠骨髓间质干细胞分化为神经元样细胞的可能性。方法 从大鼠骨髓中分离培养间质干细胞并传至第 4代 ,诱导前 2 4h加 10 ng/ m l碱性成纤维生长因子 ( b FGF)入培养液中以促分裂 ,再以含维生素 A酸、锌及大鼠损伤脊髓提取液的有血清培养液作诱导剂 ,观察细胞形态的变化 ,并采用免疫组织化学法 ,对诱导后第 12 d的细胞进行神经元特异性烯醇化酶 ( NSE)、神经丝蛋白( NF)和胶质纤维酸性蛋白 ( GFAP)表达的检测。结果 诱导后第 12 d部分细胞不仅在形态上表现为神经元样 ,而且呈 NSE及 NF阳性表达 ,GFAP阴性。结论 经维生素 A酸。

Objective To explore the feasibility of inducing rat bone mesenchymal stem cells (BMSC) to differentiate into neuron-like cells with the use of VitaminA acid, zinc and rat injured spinal cord extracts in vitro. Methods The BMSC were isolated from rat, cultured for 4 passages, and were treated with 10 ng/ml basic fibroblast growth factor (bFGF) for 24 h before induction. Then the medium was replaced by an induction media containing VitaminA acid, zinc and rat injured spinal cord extracts. The morphological changes of the cells were observed. At day 12 of induction, the cells were stained immunocytochemically with neuron-specific enolase (NSE), neurofilament (NF) and glial fibrillary acidic protein (GFAP) antibodies. Results At day 12 of induction, a certain number of BMSC became neuron-like cells and showed NSE and NF expression. But the neuron-like cells did not express GFAP. Conclusion The BMSC can be induced to differentiate into neuron-like cells with the use of VitaminA acid, zinc and rat injured spinal cord extracts.