摘要
目的 从胚胎豚鼠端脑组织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 分离胚胎豚鼠端脑组织 ,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)的表达及细胞分化后胶质纤维酸性蛋白 (GFAP)的表达。采用荧光染料Hoechst33342标记神经干细胞。结果 从胚胎豚鼠端脑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和分化成神经元和神经胶质细胞 ;荧光染料的标记效率可达 96 .7% ,细胞传代 6次后荧光亮度仍无明显衰减。结论 从胚胎豚鼠端脑分离的细胞具有明显的增殖能力及多分化潜能 ,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 fetal guinea pig were made to creat bases for the experimental study on treating sensorineural deafness by transplantation of neural stem cells. Method Neural stem cells from corpus striatum tissue of fetal guinea pig were isolated and cultured by serum-free medium containing basic fibroblastgrowth factor (bFGF) and epidermal growth factor (EGF) and identified with nestin immunocytochemical staining. The differentiated cells were checked up with glial fibrillary acidic protein (GFAP) immunocytochemical staining. The neural stem cells were marked with fluorescent dye, bisbenzimide (Hoechst 33342). Result Clone-like cluster of neural stem cells were obtained from corpus striatum tissue and the cells could be differentiated into neurons and astroeytes; the label efficiency of cells with the fluorescent dye, bisbenzimide (Hoechst 33342) was 96.7% and no ettennuation of fuorochrome brightness was observed after 6 generations in culture. Conclusion The cells from corpus striatum tissue of fetal guinea pig have the potential of proliferation and differentiation ability and the neural stem cells can be efficiently labelled with fluorescent dye. They can be used as donor cells to experimental research on treating neurosensory deafness with transplantation of neural stem cells.
出处
《中国耳鼻咽喉颅底外科杂志》
CAS
2004年第2期69-71,共3页
Chinese Journal of Otorhinolaryngology-skull Base Surgery
关键词
神经干细胞
细胞
培养
细胞标记
neural stem cells
cell culture
guinea pigs
bisbenzimide
cell label
