新生豚鼠神经干细胞的分离培养、鉴定及标记  

Isolation,culture and label of neural stem cells from newborn fetal guinea pig

作　　者：王英[1] 董明敏[1] 杜英[2] 高嵩涛[3] 

机构地区：[1]郑州大学第一附属医院耳鼻喉科,河南郑州450052 [2]郑州大学基础医学院微生物与免疫教研室,河南郑州450052 [3]中南大学湘雅医院,湖南长沙410008

出　　处：《中国现代医学杂志》2006年第14期2112-2114,共3页China Journal of Modern Medicine

基　　金：国家211工程资助项目

摘　　要：目的从新生豚鼠海马中分离培养神经干细胞,为神经干细胞移植治疗感音神经性耳聋的实验研究创造条件。方法分离新生豚鼠海马组织,用含碱性成纤维生长因子(bFGF)和表皮生长因子(EGF)无血清培养技术培养神经干细胞,免疫组化技术检测其巢蛋白(Nestin)的表达。采用荧光染料DAPI标记神经干细胞。结果从新生豚鼠海马组织分离出的细胞中可获得呈集落样生长的神经干细胞团,并能表达Nestin;荧光染料的标记效率可达93.4%,细胞传代8次后荧光亮度仍无明显衰减。结论从新生豚鼠海马分离的细胞具有明显的增殖能力,荧光染料的标记可获得较高的标记效率,可作为神经干细胞移植治疗感音神经性耳聋实验研究的供体细胞。[Objective] Isolation, culture and label of neural stem cells from newborn fetal guinea pig were made to creat bases for the experimental study on treating sensorineural deafness by transplantation of neural stem ceils, [Method] Neural stem ceils from the hippocampi tissue of fetal guinea pig were isolated and cultured by serumfree medium containing basic fibroblast growth factor（bFGE） and epidermal growth factor（EGF） and identified with nestin immunocytochemical staining. The neural stem cells were marked with fluorescent dye DAPI. [Result] Clone-like cluster of neural stem cells were obtained from hippocampi tissue and the cells could expressed the nestin; the lable efficiency of cells with the fluorescent dye, DAPI was 93.4% and no attermuation of fuorochrome brightness as observed after 8 generations in culture. [Conclusion] The cells from hippocampi tissue of newborn fetal guinea pig have the potential of proliferation ability and the neural stem cells can be efficient/y labbled with fluorescent dye. They can be used as donor cells to experimental research on treating neurosensory deafness with transplantation of neural stem ceils.

关 键 词：神经干细胞 细胞培养 细胞标记 

分 类 号：R322.85[医药卫生—人体解剖和组织胚胎学] R329.2[医药卫生—基础医学]