成年 SD 大鼠海马齿状回神经干细胞分离培养和鉴定的改良

目的：改良成年SD大鼠神经干细胞分离、培养及鉴定方法，观察神经干细胞的生长、增殖及分化特点，为后续实验提供细胞。方法从成年SD大鼠分离出完整海马齿状回，采用机械吹打法获得原代细胞，用accutase消化传代，利用cck-8法检测神经干细胞的增殖情况，利用多重免疫荧光细胞化学方法鉴定神经干细胞及其分化细胞。结果机械吹打法可高效获得原代神经干细胞，accutase消化传代更有利于神经干细胞的传代培养，cck-8法简单高效的测定了神经干细胞的增殖，多重免疫荧光创新性的动态展示了神经干细胞经诱导分化后的分化过程。结论改良后的方法可更简单高效的获得和培养出大量细胞，经多重免疫荧光鉴定所分离和培养的细胞是神经干细胞。

Improvement of isolation,culture and identification of neural stem cells from adult SD rat Hippocampus den-tate gyrus

Objective To improve method for isolating ,culturing and identifying neural stem cells （NSCs）derived from the hippocampus dentate gyrus in adult SD rat ,observe characteristics of growth ,multiplication and differentiation ,and prepare NSCs for subsequent experiments.Methods The whole hippocampus dentate gyrus was separated from adult SD rat. Primary cells were acquired by making use of mechanical blow. The neural cell pellet were passaged by accutase digestion method ,pro-liferation of NSCs could be determined by cck-8 method. Cultured and differentiated cells were identified with multiple immuno-fluorescence cytochemistry method. Results Primary neural stem cells could be efficiently obtained by mechanical blow ,ac-cutase digestive method was more conducive to the neural stem cells in the process of subculture ,the CCK-8 method was simple and efficient to determine the proliferation of neural stem cells ,multiple immunofluorescence innovatively dynamically displayed the differentiation process of neural stem cells after being induced.Conclusion The improved method is more simple and effi-cient in obtaining and culturing a large number of cells. The isolated and cultured cells are determined to be neural stem cells by multiple immunofluorescence.