大鼠视网膜共培养诱导神经干细胞分化为视网膜神经节细胞

目的:研究新生大鼠视网膜组织与胚胎上丘源神经干细胞共培养对干细胞分化的影响.方法:采用机械分离、无血清培养基选择培养的方法从孕14 d的胚鼠上丘中获取神经干细胞并传代培养.使用组织-细胞共培养系统将出生4 d的乳鼠视网膜组织与上丘源神经干细胞共培养,观察干细胞分化过程,对分化细胞进行鉴定,并与干细胞的自然分化过程进行比较.结果:从胚鼠上丘中获得的细胞在无血清培养条件下聚集呈球形悬浮生长,可以持续传代培养,具有多向分化能力,经免疫荧光鉴定证实为神经干细胞.新生大鼠视网膜组织与胚胎上丘源神经干细胞共培养促进干细胞的分化,并诱导分化出Thy1.1阳性细胞.结论:从胚胎大鼠上丘可以分离培养神经干细胞,与新生鼠视网膜组织共培养可以诱导上丘源神经干细胞分化出视网膜神经节细胞.

Retina co-culture induces neural stem cells proliferation into retinal ganglion cells

Objective:To study the effects of co-culture of superior colliculus (SC) derived neural stem cells (NSCs) and retina from newborn rat on the differentiation of SC derived NSCs. Methods: NSCs were mechanically separated from SC of 14-day embryo of SD rats. SC derived NSCs were sub-cultured in a serum-free medium using selective culture. Retinal tissue from 4 d newborn rat was co-cultured with SC derived NSCs in a tissue-cell co-culture system. The differentiation of NSCs was observed dynamically and the differentiated cells were identified by immunofluorescence technique. The natural differentiation process and co-culture differentiation process of SC derived NSCs were compared. Results: Cells from SC of the rat embryo formed into masses in a suspension state under serum-free condition. The cells were sub-cultured and had multilineage potential,and they were identified as NSCs by immunofluorescence technique. Co-culture with retinal tissue from newborn rat promoted the differentiation of NSCs into Thy1.1 positive cells. Conclusion: Embryonic SC derived NSCs can be acquired by mechanical separation and serum-free culture. Co-culture with retinal tissue from newborn rat can induce differentiation of NSCs into retinal ganglion cells.