神经膜细胞促进神经干细胞存活及分化的实验研究  

Research of Schwann cells on promoting survival and differentiation of neural stem cell in vitro

作　　者：林旭妍[1] 林绿标[2] 许益民[2] 肖哲[2] 

机构地区：[1]汕头大学医学院第一附属医院眼科,广东省515041 [2]汕头大学医学院第一附属医院神经外科,广东省515041

出　　处：《中华脑科疾病与康复杂志（电子版）》2012年第5期24-27,共4页Chinese Journal of Brain Diseases and Rehabilitation（Electronic Edition）

基　　金：广东省医学科研基金资助项目(A2010395)

摘　　要：目的 通过神经膜细胞与神经干细胞的共培养,观察神经膜细胞是否能为神经干细胞的生存和分化提供一个适宜的环境。方法 神经膜细胞取自新生1d大鼠的坐骨神经,神经干细胞取自孕14～ 16 d大鼠胎鼠的前脑。神经干细胞用胎牛血清进行诱导分化。神经膜细胞及神经干细胞分别用相应的免疫组化方法进行鉴定。纯化后的神经膜细胞和神经干细胞在无血清或表皮生长因子(EGF)的基础培养基中进行共培养实验。结果 神经膜细胞呈梭形,贴壁生长,增殖十分活跃。免疫组化检测,神经膜细胞呈S-100抗原阳性。神经干细胞在含EGF的培养基中也增殖活跃,许多细胞聚集在一起形成球状,称克隆球。当培养液换成含10％胎牛血清(不含EGF)的培养基后,这些克隆球开始贴壁并分化。免疫组织化学检测,神经干细胞呈nestin抗原阳性,而分化细胞中,部分细胞呈neurofilament阳性,部分呈胶质纤维酸性蛋白(GFAP)阳性。实验组中,与神经膜细胞共培养的神经干细胞仍能存活并分化,而对照组中的克隆球逐渐分解坏死。结论 神经膜细胞在培养状态下能为神经干细胞的存活及分化提供一个适宜的环境。Objective To determine whether Schwann cells( SCs) could provide a suitable environment for the survival and differentiation of neural stem cells by the culture of SCs and neural stem cells( NSCs). Methods SCs were cultured from the sciatic nerves of 1 day old postnatal rats,and NSCs were isolated from neocortical tissue of 14 to 16 days embryonic rat. NSCs were induced and differentiated by fetal calf serum. SCs and NSCs were identified by relevant immunohistochemistry. Then SCs and NSCs were cocultured in medium without serum or epidermal growth factor( EGF). Results Shuttle-like SC cells proliferated extensively and adhered to the dishes. The cells were S-100 positive. NSCs proliferated extensively and formed neural sphere in medium supplemented with EGF. After EGF was replaced to 10%fetal calf serum,neural sphere were seen attaching to the bottom and differentiation. NSCs were nestin positive and the differentiated cells were neurofilament positive or GFAP positive. NSCs were survival and differentiated when co-culture with SCs,whereas neural spheres in control groups deteriorated daily.Conclusion SCs can offer suitable environment for the survival and differentiation of NSCs in vitro.

关 键 词：许旺细胞 干细胞 细胞存活 细胞分化 体外研究 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]