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目的:建立永生化人晶状体上皮细胞(Human lens epithelial cell,HLEC)培养、冻存与复苏技术。方法:传代培养人晶状体上皮细胞系B-3,并进行细胞冻存与复苏,台盼蓝染色法判断细胞活力。结果:接种后的HLEC在短时间内即可贴壁及延伸,12～24 h后大部分细胞贴壁生长,具有上皮细胞的形态特点,约3～4 d细胞基本融合。冻存后复苏的HLEC活力超过90%,保持正常的细胞形态,并可长期传代培养。结论:成功建立永生化HLEC培养、冻存与复苏技术,为白内障相关研究提供理想的细胞材料。
Objective: To establish immortalized human lens epithelial cell (HLEC) culture, cryopreservation and resuscitation techniques. Methods: The human lens epithelial cell line B-3 was subcultured and the cells were cryopreserved and resuscitated. Trypan blue staining was used to determine cell viability. Results: The inoculated HLEC could adhere and extend in a short period of time. Most of the cells grew adherently after 12 to 24 hours and had the morphological characteristics of epithelial cells. The cells were basically fused in about 3-4 days. HLEC vitality after cryopreservation resumed more than 90%, maintain normal cell morphology, and long-term subculture. Conclusion: The technique of immortalized HLEC culture, cryopreservation and resuscitation was successfully established to provide ideal cell material for cataract related research.