胚胎小鼠脊髓源性与纹状体源性神经干细胞的培养及分化特点

目的 探讨小鼠脊髓源性神经干细胞与纹状体源性神经干细胞的分离培养方法 及增殖特点,比较两种来源的神经干细胞发育时期上的异同,寻找更有利于脊髓损伤修复的种子细胞.方法 利用显微解剖、无血清培养和单细胞克隆技术在孕14 d小鼠的胎鼠的脊髓及纹状体中分离培养具有单细胞克隆能力的细胞,免疫荧光染色检测克隆细胞的神经巢蛋白(nestin)抗原和诱导分化后特异性成熟神经细胞抗原的表达,并比较两种来源的干细胞在培养及分化方向上的异同点.结果从胎鼠的脊髓和纹状体中成功分离出神经干细胞.两种来源的干细胞均具有连续克隆能力可传代培养,表达nestin.脊髓血清诱导分化后脊髓源性神经干细胞β-tubulinⅢ阳性细胞(13.5±0.8)较纹状体源性神经干细胞(17.4±1.1)减少,而nestin、GFAP阳性细胞明显增多(45.7±0.3vs 39.2±1.2;25.2±1.3 vs 18.8±0.9),差异均有统计学意义(P<0.05). 结论 依据细胞增殖特点和分化结果的区别,证实纹状体源性神经干细胞更适合用于移植修复脊髓损伤.

In vitro culture and differentiation characteristics of neural stem cells derived from embryonic mouse spinal cord and striatum

Objective To investigate the isolation and culture of the neural stem cells (NSCs) derived from the embryonic mouse spinal cord and striatum, and observe the differences in their proliferation and differentiation characteristics in vitro, thereby providing evidences for identifying more suitable seed cells for repainng spinal cord injuries. Methods The spinal cord and striatum of 14-day-old embryonic mice were dissected for primary NSC culture. After 4 passages, the cells were examined for the expression ofnestin, β-tubulin Ⅲ, glial fibrillary acidic protein (GFAP) and MAG using immunohistochemistry. Results The NSCs derived from the spinal cord and the striatum both possessed in vitro proliferation capacity and expressed nestin antigen. After induced differentiation, the NSCs expressed specific antigens of neurons, astrocytes and oligodendrocytes. Conclusion Multipotent NSCs can be obtained from embryonic mouse spinal cord and striatum through different in vitro culture methods under different conditions. According to the differenees between the two kinds of NSCs in culture situation and differentiation, the striatum derived NSC is suitable for transplantation to repair spinal cord injury.