摘要
目的建立分离培养及鉴定小鼠肠神经嵴干细胞(gut neural crest stem cells,GNCSCs)的方法,为临床应用奠定基础。方法分离胚鼠肠管,消化后接种于添加N2、B27和碱性成纤维细胞生长因子的DMEM/F12培养基,贴壁培养;连续传代后用血清促进GNCSCs分化;最后用免疫细胞化学方法染色鉴定。结果分离培养的细胞聚集生长,形成的克隆球可以传代;克隆球p75染色阳性;分化后的细胞分别表达肠神经元、神经胶质细胞和平滑肌细胞特异性抗原。结论分离培养的克隆球具有自我更新和多向分化的能力,表达肠神经嵴干细胞的特异性抗原p75,是肠神经嵴干细胞。
Objective To establish a satisfactory method of isolating, culturing and determining gut neural crest stem cells, which may provide theoretical basis for clinical application. Methods The guts of embryonic mice removed and dissociated were plated into serum-free DMEM/F12 medium. The mitogen-free DMEM/F12 medium supplementd with 10% fetal bovine serum was used to induce differentiation of GNCSCs. Neurospheres and their derivations were determined with immunocytochemical and immunofluorescent staning. Results Neurospheres were generated in the simplified serum-free medium. The staining results showed that enteric neurospheres were GNCSCs and could differentiate into neurons, glial cells and smooth muscle cells by serum-induction. Conclusion GNCSCs have the capacity of self-renewal and proliferation, and give rise to neurons, glial cells and myofibroblasts.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2007年第5期595-597,共3页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金资助项目(No.30571932)
关键词
肠神经嵴干细胞
胚胎小鼠
细胞培养
细胞分化
先天性巨结肠症
gut neural crest stem cell
embryonic mouse
cell culture
cell differentiation
hirschsprung disease
