骨髓基质细胞转化为神经细胞的实验研究(英文)

Transdifferentiation of bone marrow stromal cells into neurons in vitro

背景研究发现骨髓基质细胞移植后能够游走到全身不同部位,增生转化为相应细胞,能不同程度恢复损伤器官功能。以证实骨髓基质细胞可在体外转化成中胚层和内胚层细胞。目的探讨骨髓基质细胞体外转化为神经细胞的可能性。设计设立对照的重复测量设计。地点和对象哈尔滨医科大学附属第一医院骨科。健康雄性SD大鼠,120～150g,清洁级,由哈尔滨医科大学附属第一医院动物室提供。干预骨髓基质细胞原代培养、传代后使用3种不同方法诱导。行大体观察,免疫组织化学及细胞电生理检查和长期培养研究。主要结局观察指标骨髓基质细胞诱导、细胞免疫组化和细胞电生理检测结果。结果第1诱导方案加入诱导剂1h50%转化成神经细胞,90min70%～80%转化为神经细胞。第2诱导方案加入诱导剂24h10%转化成神经细胞。第3诱导方案BMSC40%加入,24h90%转化成神经细胞,胞体小突触短。BMSC80%时加入,转化的神经细胞呈巢状排列占40%。骨髓基质细胞体外可诱导为突触明确的神经细胞(诱导率>80%);诱导后细胞表面有神经特异性抗原NSE,GFAP表达;同时具有早期神经细胞电流特性。结论骨髓基质细胞可横向转化为早期神经细胞。

BACKGROUND:As discovered by the researches,bone marrow stromal cells(BMSCs) could travel to different parts of the body after transplantation,proliferate and transdifferentiate into corresponding cells to recover to some extent the functions of injured organs.It has been proved that BMSCs could transdifferentiate into mesoblast and endoblast cells in vitro.OBJECTIVE:To investigate the possibility of the transdifferentiation of BMSCs into neurons.DESIGN:A controlled repeated measurement design was conducted.SETTING and PARTICIPANTS:Study was conducted in the Department of Orthopaedics,the First Affiliated Hospital of Harbin Medical University.Subjects were healthy male SD rats with a body mass from 120 g to 150 g in clean grade obtained from the Laboratory Animal Department,the First Affiliated Hospital of Harbin Medical University.INTERVENTIONS:Three different schemes were used to induce the BMSCs after primary and passage culture.General observation,immunocytochemistry and electrophysiology examinations as well as long term culture study were conducted.MAIN OUTCOME MEASURES:The results of BMSC induction,immunocytochemistry and electrophysiology.RESULTS:One hour after adding inducer in the first induction scheme,there were 50%of BMSCs transdifferentiating into neurons and 70%-80%into neurons at 90 minutes.At 24 hours after adding inducer in the second induction scheme,there were 10%of BMSCs transdifferentiating into neurons.40%of BMSCs were adding in the third scheme,there were 90%of BMSCs transdifferentiating into neurons at 24 hours with small soma and short synapse;when 80%of BMSCs were adding,40%of the transdifferetiated neurons were in a nest arrangement.BMSCs could be transdifferentiated into neurons with clear synapse(induction rate >80%),which expressed specific markers of neuron,neuro specific enolase and glial fibrillary acidic protein,and also had the current characteristics of the early neurons at the same time.CONCLUSION:It is possible that BMSCs can be induced into early neurons.