大鼠背根神经节源干细胞分离培养和鉴定

目的：探讨大鼠背根节（DRG）内是否存在神经干细胞。方法：取胚胎大鼠DRG进行原代细胞培养,观察其增殖与分化情况,并行P75BrdU/Nestin,GFAP,NF200免疫荧光组织化学鉴定。结果：细胞培养获得大量能够分裂增殖且呈巢状悬浮生长的干细胞团,呈P75BrdU/Nestin免疫荧光组织化学阳性反应;培养第三代细胞加入胎牛血清10d后,相差显微镜下可见部分细胞伸出多个细长突起,且免疫组化呈NF200阳性;部分细胞具有扁平多突起的或双极细胞形态,免疫组化呈GFAP染色阳性。细胞球经NGF培养诱导1d左右有些神经元样的细胞开始迁出,随着时间增长迁出的细胞越来越多,并且细胞和细胞之间发生联系,形成交错的复杂网络,迁出的细胞经免疫组化鉴定几乎都是呈NF200免疫染色阳性。结论：大鼠DRG内存在有神经干细胞,这些细胞可以分化为神经元和神经胶质细胞。

Culture and identification of neural stem cells from rat dorsal ganglia

Objective:To investigate whether there exist neural stem cells in rat dorsal ganglia(DRG).Method:Cells isolated from fetal rat DRG were respectively cultured and observed in their proliferation and differentiation.The cellular multiplication was observed by immunocytochemical fluorescen with nestin and co-label with Brdu.The differentiated cells were detected with neurofilament protein 200(NF200) and Glial Fibrillary Acidic Protein(GFAP) immunocytochemical fluorescent staining respectively.Result:Nest-like clusters of neural stem cells were obtained in suspension and the cells could devide or proliferate and showed Nestin/Brdu positive.Some of the third passaged cells protruded several elongate processes and expressed the NF200 in the presence of fetal bovine serum,while some cells were flat or dipolar and labled with GFAP.There were some cells migrated from the neurosphere after adding NGF two days later,and the time was longer,the cells were more,those cells contacted with each other and form a complicated net,and furthermore,the cells could be labled with NF200.Conclusion:There are neural stem cells in rat DRG,which possess the abilities of division,multiplication and self-renew,and they can differentiate into neuron and glial cells.