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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。
OBJECTIVE: To observe the effect of LipofectamineTM 2000 on human corneal stromal cells and to explore its feasibility and safety scope for human corneal stromal cells. Methods: Human corneal stromal cells were cultured in vitro. The cell lines 3-5 were used for the experiment. The effects of different concentrations and time of liposomes on the proliferation rate of human corneal stromal cells were detected by MTT assay. The effect on the survival rate was detected by trypan blue staining. Results: The influence of liposomes on human corneal stromal cells was related to the concentration and the action time. Concentration higher than a certain level can cause cell proliferation rate and survival rate decreased at the same concentration, the longer the role of the more obvious decline. A concentration of less than 40mg / L for 24 hours will not have a significant effect on cell proliferation rate and survival rate. Conclusion: LipofectamineTM2000 does not cause cytotoxicity in a certain range and is expected to play an important role in the gene therapy of corneal stromal cells.