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目的:探讨人、鼠视网膜Muller细胞体外培养的方法,并研究视网膜Muller细胞谷氨酸转运体GLAST(L-谷氨酸/L-门冬氨酸转运体)的表达与分布。方法:采用视网膜组织块的方法培养引产胎儿及产后3 d(P3)乳鼠视网膜Muller细胞,振荡法分离纯化细胞,观察细胞形态和生长特性,并对Muller细胞进行免疫细胞化学荧光染色。结果:采用组织块培养的Muller细胞,约需15~20 d细胞基本融合,振荡法分离的细胞纯度高。培养的细胞神经胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、波形蛋白(Vimentin)及其谷氨酸转运体GLAST抗体染色阳性,GLAST主要分布在Muller细胞膜及突起上。结论:用组织培养的方法可成功培养出人、鼠视网膜Muller细胞,Muller细胞膜上有广泛的谷氨酸转运体GLAST的表达及分布。
OBJECTIVE: To investigate the in vitro culture of human and mouse retinal Muller cells and investigate the expression and distribution of GLAST (L-glutamate / L-aspartate transporter) in retinal Muller cells. Methods: Retinal Muller cells were induced in fetus of fetus and postnatal day 3 (P3). Retinal Muller cells were isolated and purified by oscillatory method. Cell morphology and growth characteristics were observed, and Muller cells were immunocytochemically stained. Results: The Muller cells cultured in tissue culture blocks needed about 15-20 days to fuse with each other. The cells isolated by shaking method had high purity. Cultured glial fibrillary acidic protein (GFAP), vimentin (Vimentin) and its glutamate transporter GLAST antibody staining, GLAST mainly in Muller membrane and protrusions. CONCLUSION: Human and mouse retinal Muller cells can be successfully cultured by tissue culture method. The expression and distribution of glutamate transporter GLAST on Muller cell membrane are abundant.