目的探索体外培养兔视网膜Mller细胞的有效方法。方法采用酶消化法从乳兔视网膜获取Mller细胞,通过振荡和反复洗涤使之纯化。采用免疫组织化学技术和透射电镜对传代Mller细胞进行鉴定。结果培养24h后即有部分细胞贴壁生长,72h后贴壁细胞进一步增多。通过振荡吹打可使附着于Mller细胞上的神经元脱落。20～25d后细胞接近融合,呈三角形或不规则形。传代后细胞经1周达到融合。培养细胞GFAP阳性率在95%以上。电镜观察显示:细胞内含有线粒体、粗面内质网、核糖体及8～10nm的中间丝。结论利用酶消化法可成功分离兔视网膜Mller细胞,通过振荡和吹打洗涤,可使之纯化。 Objective To explore an effective method of culturing rabbit retinal Mller cells in vitro. Methods Mller cells were obtained from the rabbit retina by enzyme digestion and purified by shaking and washing repeatedly. The passage Mller cells were identified by immunohistochemistry and transmission electron microscopy. Results After culturing for 24h, some cells adherently grew, and the adherent cells further increased after 72 hours. Neurons attached to Mller cells shed by oscillating blow. After 20 ~ 25 days, the cells were close to fusion and showed triangular or irregular shape. After passage the cells reached confluence by 1 week. The positive rate of GFAP in cultured cells was over 95%. Electron microscopy showed that the cells contained mitochondria, rough endoplasmic reticulum, ribosomes and 8 ~ 10 nm intermediate filaments. Conclusion Rabbit retinal Mller cells can be successfully isolated by enzymatic digestion, and purified by shaking and washing with blower.