组织-细胞共培养诱导神经干细胞定向分化的研究  

The experimental study on the directional differentiation of neural stem cells induced by tissue-cell co-culture

作　　者：冯东福[1] 卢亦成[2] 楼美清[2] 刘勇[1] 陈二涛[1] 汪洋[3] 

机构地区：[1]上海交通大学医学院附属第三人民医院神经外科 ,201900 [2]第二军医大学附属长征医院神经外科 [3]复旦大学上海医学院解剖与组织胚胎学系

出　　处：《中华实验外科杂志》2008年第2期236-238,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30271393）;上海市教委自然科学基金资助项目（04BB23、07ZZ42）

摘　　要：目的探讨大鼠上丘组织-神经干细胞（NSCs）共培养对细胞分化的影响。方法采用元血清选择培养的方法获得上丘源NSCs，单细胞克隆传代培养并进行形态学观察。使用组织-细胞共培养系统将不同年龄大鼠上丘组织与NSCs共培养，观察NSCs分化过程。对分化细胞进行鉴定，并与NSCs的自然分化过程进行比较。结果从胚鼠上丘中获得的细胞呈球形悬浮生长，可形成单细胞克隆，具有多向分化能力，经免疫荧光鉴定证实为NSCs。新生大鼠上丘组织与NSCs共培养可以促进NSCs的分化，并诱导分化出Thy1.1阳性的视网膜神经节细胞，阳性细胞比率为19．97％（273／1094）。而神NSCs的自然分化或与成年大鼠上丘组织共培养后均未发现Thy1.1阳性细胞。结论从胚胎大鼠上丘可以分离培养出NSCs，与新生大鼠的上丘组织共培养可以诱导NSCs分化为视网膜神经节细胞。Objective To investigate the effects of co-culture of neural stem cells （NSCs） with superior colliculus （SC） explants of rat on the differentiation of the NSCs. Methods NSCs were mechanically separated from the SC of the embryo of 14-day pregnant SD rats and sub-cultured in a serum-free medium by using selective culture. Single-cell-clones were isolated and passaged, and morphologic change of clones were observed. NSCs were co-cultured with SC explants from different age rats in a tlssue-cell coculture system. The differentiation state of NSCs was observed dynamically, and by using the immunofluorescence technique, the differentiated cells were identified. The natural differentiation process of SC derived NSCs was also compared with co-culture differentiation process of NSCs. Results Cells from SC of the rat embryo gradually scattered cell masses in a suspension state under the serum-free condition. Single cell clone was cultured arid had multi-directional differentiation potential. These SC derived cells were identified as NSCs by using the immunofluorescence technique. Co-culture with SC explants from newborn rats could improve the differentiation of NSCs and induce differentiation of NSCs into Thyl. 1 positive cells. The positive rate was 19.97% （273/1094）. No Thyl. 1 positive cells were found in natural differentiation of NSCs or Co-culture with SC explants from adult rats. Conclusion NSCs can be acquired from embryonic anterior colliculus in rats. Co-culture with SC explants from newborn rats could induce differentiation of NSCs into retinal ganglion cells.

关 键 词：干细胞 组织培养 视网膜神经节细胞 分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R996[医药卫生—基础医学]