Abstract:AIM:To investigate the expression level of rabbit corneal epithelial cells'' basic fibroblast growth factor (bFGF) in vitrounder the condition of using different concentration of human amniotic homogenate supernatant, and to discuss the function of human amniotic homogenate supernatant in the process of corneal epithelial repair.METHODS:The rabbit corneal epithelial cells were cultured in vitroand they were divided into several groups. Then the experiment group was treated in DMEM culture liquid of 100mL/L fetal blood serum mixed with different concentration of human amniotic homogenate supernatant (the amniotic homogenate protein concentration was 40,80,160μg/mL respectively), with the control group treated in DMEM culture liquid of 100mL/L fetal blood serum only. By using the method of methyl thiazolyl tetrazolium(MTT) colorimetry, we aim to investigate the impact of human amniotic homogenate supernatant on rabbit corneal epithelial cells proliferation at different times (24, 48, 96 hours). At the same time, we tested the bFGF-mRNA expression level in each group by RT-PCR.RESULTS:Rabbit corneal epithelial cells proliferated at different concentrations of human amniotic homogenate supernatant (40, 80, 160μg/mL), the amount of corneal epithelial cells in each experimental group was increased significantly (0.185±0.010, 0.318±0.015, 0.501±0.014 respectively, P<0.05). As the culture time prolonged, the difference became more apparent; There was no significant difference about the expression of bFGF-mRNA between the control(0.708±0.013) and the amniotic homogenate supernatant with the concentration of 40μg/mL and 80μg/mL (0.750±0.007, 0.785±0.006 respectively), but the differences about the expression of bFGF-mRNA between the 160μg/mL group(1.013±0.120) and the 40, 80μg/mL, control group were significant respectively.CONCLUSION:Human amniotic homogenate supernatant can significantly stimulate the proliferation of rabbit corneal epithelial cells and its effect shows dose-dependant; Human amniotic homogenate supernatant could promote the expression of the bFGF-mRNA in rabbit corneal epithelium cells in vitro, which is concentration related.
