兔骨髓间充质干细胞向神经元的诱导分化

目的 :探讨兔骨髓间充质干细胞 (bonemarrowmesenchymalstemcells ,BM -MSCs)向神经元定向诱导分化及分化前后体外培养的条件。方法 :无菌条件下收集兔股骨骨髓 ,用相对密度为 1 0 77的淋巴细胞分离液分离出白膜层细胞群 ,贴壁法筛检其中的MSCs ,用DMEM H条件培养基进行原代及传代培养。分别取 5和 10代的MSCs向神经元定向诱导。诱导后更换为神经培养基继续培养 ,并在 1～ 2 0d用免疫细胞化学方法检测分化细胞的特异性表面标志。结果 :兔骨髓MSCs在DMEM H条件培养基中建立高纯度的细胞培养 ,能够稳定地连续传代达 10代以上。不同代数的MSCs诱导后均表达巢蛋白 (nestin)和神经特异性烯醇化酶 (neuron specificenolase ,NSE)。诱导后NSCs在神经培养基中继续存活 2 0d以上 ,但未见扩增。结论 :兔骨髓MSCs能够诱导表达神经元特异性标志 ,且诱导前后的MSCs均可在体外培养条件下存活。

Neuron-oriented differentiation from rabbit bone marrow mesenchymal stem cells

Objective: To explore the induction from rabbit bone marrow mesenchymal stem cells(BM-MSCs) into neurons and the culture in vitro pre-and post-induction.Methods: Rabbit bone marrow samples were obtained from femoral cavities,from which mononuclear cells layers were isolated by lymphocyte seperation medium,and MSCs were then selected to initiate cell culture as the adherent component.MSCs grew in DMEM-H conditional medium and passaged continually.The 5 and 10 passages MSCs were induced to differentiate to neuron direction.Induced cells were cultivated consequently with neuron-specific medium,and molecular markers were detected immunocytochemically during 1 to 20 days respectively.Results: Highly purified rabbit BM-MSCs culture were established in the DMEM-H conditional medium and passaged steadily over 10.MSCs of different passages expressed nestin and neuron specificenolase(NSE).Induced cells survived in neuron-specific medium no less than 20 days,but observed no proliferation.Conclusion: Rabbit BM-MSCs can be induced to express neuron-specific markers and can grow pre-and post-induction in vitro.