躯干神经嵴干细胞体外分离培养及其增殖、分化的实验研究  

Culture,proliferation,and differentiation of trunk neural crest stem cells in early embryos of rats:an in vitro experiment

作　　者：卢静[1] 孙晋浩[1] 张静[1] 丛丽[1] 綦晓霞[1] 庄园 杨洁[1] 李振华[1] 

机构地区：[1]山东大学医学院解剖学教研室 [2]组织与胚胎教研室,济南250012

出　　处：《山东大学学报（医学版）》2009年第4期46-49,共4页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金资助课题(Y2007C031);山东省自然科学基金资助课题(Y2007C003)

摘　　要：目的体外分离培养鉴定躯干神经嵴干细胞,探讨其生物学特性,为研究周围神经再生修复提供新的细胞来源。方法取大鼠胚胎躯干部神经管,组织块法培养获取躯干神经嵴干细胞,通过形态学及免疫细胞化学观察其细胞生长、增殖、分化等特性。结果原代培养的躯干神经嵴干细胞呈梭形,其特异性标记物nestin和p75表达阳性,并有较高的增殖活性。传代培养后,在含血清培养基中,其分化细胞呈GFAP、MAP-2、α-SMA阳性。结论成功地培养了躯干神经嵴干细胞,细胞表现出多潜能干细胞特性,为研究周围神经组织工程种子细胞来源提供实验依据。Objective To establish a method for in vitro culture of tnmk neural crest stem cells and to explore their biological properties, so as to find a new treatment for peripheral nervous system diseases. Methods A neural tube towards the candal- most 10 somites obtained from fetal rats at E10.5 was isolated and explanted in a plate coated with fibmnectin for 2 days. Truncal neural crest stem cells were collected after being trypsinased. Their growth proliferation and differentiation were observed by an inverted phase contrast microscope and immunocytochemical technique. Results Primary culture of truncal neural crest stem cells with fibmblast-like appearance demonstrated characteristics of neural stem cells, such as clonality and pluripotency. Immu- nocytochemical staining showed that the cells expressed nestin and p75. Truncal neural crest stem cells could differentiate into neurons, Schwann cells and smooth muscle-like cells in the serum-containing medium. Conclusion Tnmk neural crest stem cells were successhtlly cultured in vitro, which provides an experimental basis for further research of tissue engineering of candi- date cells.

关 键 词：神经嵴 干细胞 细胞培养 细胞分化 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学]