盖玻片辅助人晶状体上皮细胞原代培养法

目的：建立人晶状体上皮细胞原代培养的简便方法并比较不同来源人品状体上皮细胞的生物学特性。方法：取胎龄20周合法引产胚胎眼晶状体囊膜、中山眼科中心眼库眼晶状体囊膜和白内障患者术中撕取的前囊膜，分别在培养皿中铺平，加10乩10％DMEM培养液润湿后加盖盖玻片防止卷曲并促进粘贴．添加培养液浸没盖玻片，37℃培养。同时取相同来源的囊膜按照组织块法培养。观察细胞增殖情况并比较原代人晶状体上皮细胞与人晶状体上皮细胞系SRA01／0413晶体蛋白的表达差异。结果：在盖玻片辅助下，胚胎眼晶状体囊膜第2天即可见明显的增殖细胞由囊膜缘长出，眼库眼囊膜和白内障患者术中撕取的囊膜在3～4d的潜伏期后亦可见增殖细胞长出；组织块法培养出现部分组织块漂浮，且胚胎眼囊膜潜伏期延长至3-4d，眼库眼囊膜和白内障患者晶状体囊膜潜伏期延长至4-5d。结论：盖玻片辅助的改良组织块培养法能尽快获得体外培养的原代晶状体上皮细胞，且操作简便，值得推广应用于品状体病的研究。

Coverslip Assisted Primary Tissue Culture for Human Lens Epithelial Cells in Vitro

PURPOSE: To set up an easy procedure of tissue culture for human lens epithelial cells in vitro and to observe the biological characteristics. METHODS: Capsules from embryo of 20 weeks, eye bank of Zhongshan Ophthalmic Centre and patients with cataract were spread on culture utensil. 10 microL of 10% DMEM medium was added and a piece of coverslip was lay to prevent crimp. Then the capsules were cultured under 37 degrees C after adding enough medium. Capsules from the same source were cultured by traditional tissue culture method. Expressions of 13 crystallin between primary tissue culture cells and SRA01/04 cell line were compared by western blotting. RESULTS: With coverslip assisted, the cells could be observed proliferated and migrated from the edge of embryo capsule 2 days later, and for capsules from eye bank and age-related cataract patients, the interval time was 3 to 4 days. By traditional tissue culture method, the interval time of embryo capsule was 3 to 4 days, and for capsules from eye bank and age-related cataract patients, the interval time was the same. And capsules floated sometimes. CONCLUSIONS: By coverslip assisted primary tissue culture human lens epithelial cells could grow faster and easier, and the method is worthy to be spread in research of lens diseases.