脂质体对人角膜基质细胞影响的实验研究(英文)

Influence on cultured human keratocytes by liposome

目的:观察脂质体LipofectamineTM2000对人角膜基质细胞的影响,探索其应用于人角膜基质细胞的可行性及安全范围。方法:体外培养人角膜基质细胞,取其第3-5代细胞鉴定后用于实验。采用MTT法检测不同浓度和时间脂质体对人角膜基质细胞增殖率的影响;采用台盼蓝染色法检测对存活率的影响。结果:脂质体对人角膜基质细胞的影响与浓度和作用时间有关。浓度高于一定水平时可引起细胞增殖率和存活率的下降,浓度相同时作用时间越长下降越明显。浓度低于40mg/L作用24h不会对细胞增殖率和存活率产生明显影响。结论:LipofectamineTM2000在一定范围内不引起细胞毒性,有望在角膜基质细胞的基因治疗中发挥重要作用。

AIM： To observe the effects on human keratocytes by cationic liposome LipofectamineTM 2000 （LF2000）, to investigate the efficiency and safe range applied in human keratocytes, and establish basis for gene therapy of human keratocytes, METHODS： Human keratocytes cultured in vivo within 3 to 5 passages were used in experiment after being identified. The effects on proliferation of cultured human keratocytes by LF2000 with different concentrations and time were evaluated By MTT; the effects of LF2000 on the survival rate and its relation with 5, 10, 20, 40, 80mg/L concentration and time were detected by trypan blue staining. RESULTS： LF2000＇s effects on human keratocytes were related with concentration and time. The cellular proliferation and survival rate declined when concentration of LF2000 was above certain level, and this effect increased as time became longer. LF2000 had no effect with concentration under 40mg/L for 24 hours. CONCLUSION： LF2000 did not cause cytotoxicity during a concentration range ＂tested＂, and it is hoped to play an important role in gene therapy of human keratocytes.