新方案诱导大鼠骨髓间充质干细胞诱导分化为神经元样细胞

目的探索体外诱导间充质干细胞分化为神经元样细胞后长期培养的可能性。方法自成年Wistar大鼠红骨髓分离培养骨髓间充质干细胞,通过克隆培养、成骨分化、成脂肪分化鉴定其特性,取第3代细胞重新接种,经b-FGF预诱导及β-巯基乙醇诱导后,换维持培养液维持培养。结果本实验分离培养的细胞克隆形成率64±1.4%,具有成骨及成脂肪分化能力。成神经元诱导分化0.5h后,多数细胞即开始呈现出神经元样外观,5h后绝大多数细胞表现出典型的神经元样外观,维持培养1周后,大多数细胞继续维持神经元样外观,72±3.6%的细胞MAP-2免疫细胞化学染色阳性。结论本实验中分离培养的细胞表现出间充质干细胞的特性,经诱导后,超过半数的细胞分化为神经元样细胞,在我们优化的培养体系中,分化的神经元样细胞可存活1周以上。

Maintaining culture following neuronal induction of rat marrow derived mesenchymal stem cells

Objective To find a protocol of sustaining culture of differentiated neuron like cells induced from marrow derived mesenchymal stem cells(MSCs).Methods MSCs were isolated from the femurs of adult Wistar rats and maintained in a-MEM supplemented with 10% Fetal Bovine Serum,the character of MSCs was identified by the generation of clone lines,osteogenic and lipogenic induction,and then a protocol of neuronal induction and maintaining culture was tested.Results Half of an hour following neuronal induction some cells showed neuron like appearance,and 5 hours later almost all cells showed typical neuronal appearance.After 7 days,most of the differentiated cells expressed the neuronal markers MAP-2.Conclusions With our induced culture protocol,MSCs can differentiate into neuron like cells and keep alive exceeding one week.