人晶状体上皮细胞原代培养方法的改进

目的 建立一种简单有效的人晶状体上皮细胞体外培养的方法.方法 用组织块培养法,采用连续花瓣状撕囊法获得人白内障患者的晶状体前囊膜,使用含有表皮生长因子的培养液进行培养,对培养的细胞进行形态学观察及鉴定;在倒置相差显微镜下观察其生长规律,应用免疫荧光组织化学法进行鉴定.结果 体外培养的原代人晶状体上皮细胞在组织块贴壁24 h后从组织块边缘长出,具有上皮细胞的形态特点.αA-晶状体蛋白抗体阳性率几乎100％,鉴定为晶状体上皮细胞.结论 采用连续花瓣状撕囊法获得的组织块较易在体外培养出入晶状体上皮细胞,可用于白内障后囊膜混浊发病机制的进一步研究.

Improvement of primary culture method for human lens epithelial cells

Objective To establish a simple and effective procedure for primary culture of human lens epithelial cells in vitro.Methods The specimens of human lens capsule from the patients with cataract were obtained with petal-like-tearing-capsule and cultured using the method of tissue explant.The growth medium was Dulbecco’s modified Eagle’s medium supplemented with 100 mL/L fetal bovine serum and 10 ng/mL recombinant rat epidermal growth factor.The morphology of cultured cells was observed under the phase contrast microscope and identified by immunocytochemistry.Results Human lens epithelial cells under primary culture in vitro grew out of the edge of tissue pieces after 24-hour incubation.The primary cultured cells maintained typical morphology of the epithelial cells.And almost 100% of the lens epithelial cells were positive for αA-crystallin antibody.Conclusion Human lens epithelial cells are easily cultured in vitro using the procedure with petal-like-tearing-capsule.This method may be used in studies on pathogenesis of cataract and posterior capsular opacification.