大鼠视网膜神经节细胞的培养

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。

Rat retinal ganglion cells in culture

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 x 10(5) cells/well) and cultured under 37 degree C 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.