小鼠视网膜干细胞的分离与鉴定  

Isolation and identification of retinal stem cells in mouse eye

作　　者：魏勇[1] 应大君[1] 朱楚洪[1] 张伟[1] 董世武[1] 

机构地区：[1]第三军医大学基础医学部解剖学教研室,重庆400038

出　　处：《第三军医大学学报》2003年第24期2161-2164,共4页Journal of Third Military Medical University

摘　　要：目的　小鼠视网膜干细胞的分离与鉴定。方法　①对胎龄 17d昆明小鼠眼睫状体部 (CMZ)细胞进行分离与体外培养 ;②通过免疫荧光方法研究视网膜干细胞的增殖及分化特性。结果　①在bFGF作用下 ,所培养细胞可形成悬浮生长的细胞团 ,经多次传代 ,细胞增殖能力未见衰减。BrdU标记结果显示悬浮细胞团主要是由分裂增殖的细胞组成。而且培养细胞可表达神经干细胞的特异性抗原Nestin及胚胎发育早期视网膜内原始细胞的特异性抗原Chx 10。②培养细胞经 5 %胎牛血清诱导可向成熟细胞分化 ,分别表达神经元与神经胶质细胞的特异性抗原NSE、GFAP。免疫荧光检测显示分化细胞可分别表达感光细胞、双极细胞以及节细胞的特异性抗原Opsin、PKC及 β tubulin。Objective To isolate the retinal stem cells from mouse eye and identify their properties. Methods The cells derived from the ciliary marginal zone(CMZ) of 17 d embryonic mouse were isolated and cultured in vitro . The proliferating capacity and differentiating properties of the cultured cells were studied by using immunofluorescence methods. Results The study showed that the isolated cells derived from CMZ cultured in serum free medium could give rise to neural spheres in the presence of bFGF. These cells could proliferate successively and generate secondary neural spheres, thus displaying potential to self renew. These neural spheres could be expanded for up to 8 passages and could express constantly the neuroectodermal marker Nestin and Chx 10, a retinal stem cell marker, showing the undifferentiated stem cells properties in vitro . Analysis of the differentiation potential of the cultured cells in vitro showed that the cultured cells derived from CMZ were multipotential. Upon withdrawal of bFGF and addition of serum, the stem cells expressed cell type specific markers corresponding to neurons and glia, such as NSE, GFAP, opsin, PKC, and β tubulin. Conclusion The CMZ in the mouse eye harbors retinal stem cells that display properties of stem cells, and these cells have the capacity to give rise to retinal neurons and glia in vitro .

关 键 词：视网膜 干细胞 分离 鉴定 

分 类 号：R322.91[医药卫生—人体解剖和组织胚胎学] R329.2[医药卫生—基础医学]