摘要
目的:采用“五步法”将鼠胚胎干细胞(embryonicstemcell,ES)诱导成为神经元和胶质细胞。方法:首先将ES细胞接种到铺有明胶的培养瓶里,在无滋养层细胞的情况下生长3d,然后将细胞悬浮培养4d成为胚胎体(,胚胎体经过无血清培养基培养6d,富集nestin阳性细胞,在有丝分裂原bFGF作用下扩增6d,最后经过烟酰胺诱导分化6d,共计经过5个步骤。结果:诱导出多量表达Tuj-1的不成熟神经元(约占分化后细胞数量的80%～90%)和表达神经胶质纤维酸性蛋白的星形胶质细胞(5%～10%),还有少量表达微管相关蛋白Ⅱ的成熟神经元(5%～10%)。结论:“五步法”诱导使胚胎干细胞在分化过程中处于不同的阶段,最终诱导出大量不成熟的神经元,少量星型胶质细胞和成熟神经元。为利用不同成熟度的分化细胞移植治疗神经系统疾病提供了充足的细胞来源。
AIM:To induce embryonic stem(ES) cells into nerve cells in vitro by 5 step method. METHODS:Firstly,ES cells were inoculated into culture bottles pretreated with gelatin,and cultured for 3 days without enriched cells,followed by suspension culture for 4 days. After the cells were transformed into embryoid body(EB),the EB was cultured in the non serum medium for 6 days to enrich nestin positive cells.The cells were amplified by cultured in basic fibroblast growth factor for 6 days.Finally,the cells were induced and differentiated by nicotinamide for 6 days. RESULTS:Among the cells that had participated in the entire induction procedure, the number of immature neurons that expressed Tui 1 was up to 80%-90%,the number of astrocytes expressing glial fibrillary acidic protein up to 5%-10%,and the number of mature neurons expressing microtubule associated protein Ⅱ(MAP Ⅱ) up to 5%-10%. CONCLUSION:Five step method makes the differentiated procedure of EB cells divided into different stages,a few astrocytes and mature neurons,which may be useful for further neurobiology study as well as potential transplantation therapy.
出处
《中国临床康复》
CSCD
2004年第10期1851-1853,共3页
Chinese Journal of Clinical Rehabilitation
基金
国家973项目"不同脑区神经干细胞的生物学特性"(001CB510104)
北京市科委科技计划项目"干细胞技术在治疗帕金森氏病中的临床应用研究"(H020220010290)
北京市卫生局重点学科项目"老年神经退行性疾病的基础与临床研究"(1998)卫科扶字12号~~
