一种急性分离视网膜细胞的实验方法  

An Optimized Isolation Protocol of Mouse Retinal Cells

作　　者：胡单萍 罗雪[1] 刘诗亮[1] 沈吟[1] 

机构地区：[1]武汉大学人民医院眼科中心,湖北武汉430060

出　　处：《武汉大学学报（医学版）》2015年第3期359-362,共4页Medical Journal of Wuhan University

基　　金：国家自然科学基金资助项目(编号:81270998;81470628)

摘　　要：目的:探讨为视网膜细胞实验提供足量且活性高的视网膜细胞的分离方法。方法:取3周的C57小鼠视网膜,用木瓜蛋白酶消化后并吹打,得到细胞悬液后用富氧的溶液灌流,待分离细胞静置沉淀后,用相差显微镜观察视网膜细胞的形态和数量。分析不同类型的视网膜细胞所需的最佳酶解时间和最佳酶浓度。结果:本实验在不同的酶解时间或酶浓度下使用木瓜蛋白酶对视网膜进行分离消化,得到的细胞数量不尽相同,通过统计得到针对每一种视网膜细胞的最优化分离条件。结论:本实验系统地研究并建立了针对分离不同视网膜细胞的最有效方法。Objective：To explore the optimized protocol of obtaining isolated mouse retinal cells.Methods：Retinas of mouse were digested by papain at different time or enzyme concentration at37℃,then were isolated under gentle mechanical force.Retinal bipolar cells,ganglion cells,Müller cells and photoreceptors are counted and the number of each type is analyzed.Results：Different types of retinal cell have their own optimized papain concentration and/or incubation time.Conclusion：We analyzed the best digestion time and enzyme concentration parameter in four types of retinal cells.Our result would benefit ophthalmology researchers in isolation retinal cells experiment in the future.

关 键 词：视网膜 木瓜蛋白酶 分离细胞 

分 类 号：R331[医药卫生—人体生理学]