[Effects of amniotic membrane on proliferation of retinal Müller cells: an in vitro experiment with rabbits].

Objective To observe the effects of amniotic membrane on the proliferation of retinal Mtiller cells.Methods(1)Human amniotic membrane was collected form normal lying woman undergoing cesarean section.Miller cells were obtained from the retina of New Zealand rabbit,cultured, and put into 96-well culture plate.Homogenate of human amniotic membrane at different concentrations: 100,200,400,and 800 μg/ml respectively were added into the culture fluid for 96 h.The proliferation rate of the Miller cells was measured with MTr method.(2)Mller cells were inoculated in 6-well culture plate with cover slips.Amniotic homogenate of different concentrations was added for 48 hours,and immunohistochemisrty was used to detect the expression of proliferating cell nuclear antigen(PCNA).(3) Amniotic homogenate conditioned culture medium on different concentrations was added into the culture fluid of Miller cells for 48 hours.The change of cell cycles was observed by flow cytometry(FCM).Results Homogenate of human amniotic membrane increased the proliferation rate of the Mller cells dese- dependently and the proliferation rate reached the peak(62.5%)when the concentration homogenate of human amniotic membrane was 800 μg/ml.FCM showed that the amniotic homogenate increased the rate of cells in S phase and decreased the rate of those in G_2/M phase and G_0/G_1 phase concentration-dependently. The proliferation rate of the Miller cells treated with the amniotic homogenate of the same concentration increased time-dependently and peaked 48 h later,and the proliferation rate 96 h later was not significantly different from that 48 h later(F=0.233,0.007,P=0.492,0.729).The positive rates of PCNA when the concentrations of the amniotic homogenate were 800 μg/ml and 400 μg/ml respectively(0.84±0.07, 0.79±0.06)were significantly higher than that of the control group(0.64±0.12).Conclusion Amniotic membrane promotes the proliferation on tretinal Mller cells dose and time-dependently in vitro.