大鼠视网膜神经节细胞的培养  

Rat retinal ganglion cells in culture

作　　者：钟一声[1] 蒋幼芹[1] 熊小玲[1] 

机构地区：[1]湖南医科大学附属第二医院眼科

出　　处：《中华眼科杂志》1999年第2期137-139,I007,共4页Chinese Journal of Ophthalmology

基　　金：国家自然科学基金

摘　　要：目的建立视网膜神经节细胞（ｒｅｔｉｎａｌｇａｎｇｌｉｏｎｃｅｌｓ，ＲＧＣｓ）的体外培养方法，为ＲＧＣｓ的体外实验研究奠定基础。方法采用胰酶消化法将１６只生后２～３天的ＳｐｒａｇｕｅＤａｗｌｅｙ大鼠视网膜制成细胞悬液后，接种于涂以鼠尾胶原的２４孔培养板，预先置入１ｃｍ×１ｃｍ的载玻片。细胞数约４×１０５个／孔，在３７℃、体积分数为５％的ＣＯ２培养箱中培养。于第１、３及５天行抗大鼠ＴＨＹ１单克隆抗体免疫细胞化学检查以鉴定ＲＧＣｓ，镜下计算每１０个高倍镜下（ｈｉｇｈｐｏｗｅｒ，ＨＰ）ＲＧＣｓ的细胞数和其轴突生长率。结果在鼠尾胶原上培养的ＲＧＣｓ生长良好，部分细胞伸出突起，且有些突起相互连接成网。培养第１天，ＲＧＣｓ数和轴突生长百分率分别为（４０１±９）个／１０ＨＰ和（２５．３４±０．７２）％，第３天为（３５１±６）个／１０ＨＰ和（３５．１６±２．２２）％，第５天为（１０９±８）个／１０ＨＰ和（６９．８４±０．９７）％。结论ＲＧＣｓ的体外培养能获成功，鼠尾胶原是ＲＧＣｓ体外生长的良好支持物。Objective To establish a culture system for retinal ganglion cells (RGCs) in order to lay a foundation for the experimental research in vitro. Methods The retinae of 16 postnatal 2～3 day Sprague Dawley rats were dissected into cell suspension with trypsin digestion. The cell suspension was implanted in 24 well culture plates with a cover slide 1 cm 2 in size and covered with murine tail collagen preplaced in each well (4×10 5 cells/well) and cultured under 37℃ in an incubator with 5% CO 2. The cells were identified by immunocytochemical method with anti rat Thy 1.1 monoclonal antibody after culture for 1, 3, 5 days, respectively, and the number of RGCs and its axon growth percentage were counted in each 10 field high power (HP, 200 x) view under light microscope. Results The RGCs cultured in murine tail collagen tissue grew very well. Some cells possessed axons and some axons connected in networks. The RGC number and its axon growth percentage were (401±9) cells/10 HP and (25.34±0.72)% in 1 day culture, (351±6) cells/10 HP and (35.16±2.22)% in 3 day culture, (109±8)cells/10 HP and (69.84±0.97)% in 5 day culture, respectively. Conclusion RGCs can be cultured successfully and the murine tail collagen tissue is a good substratum for RGC survival in vitro.

关 键 词：视网膜 神经节细胞 细胞培养 青光眼 

分 类 号：R775.02[医药卫生—眼科] R329－33[医药卫生—临床医学]