[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 Müller cells.


METHODS
(1) Human amniotic membrane was collected form normal lying woman undergoing cesarean section. Müller 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 microg/ml respectively were added into the culture fluid for 96 h. The proliferation rate of the Müller cells was measured with MTT method. (2) Müller cells were inoculated in 6-well culture plate with cover slips. Amniotic homogenate of different concentrations was added for 48 hours, and immunohistochemistry 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 Müller 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 Müller cells dose-dependently and the proliferation rate reached the peak (62.5%) when the concentration homogenate of human amniotic membrane was 800 microg/ml. FCM showed that the amniotic homogenate increased the rate of cells in S phase and decreased the rate of those in G2/M phase and G0/G1 phase concentration-dependently. The proliferation rate of the Müller 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 microg/ml and 400 microg/ml respectively (0.84+/-0.07, 0.79+/-0.06) were significantly higher than that of the control group (0.64+/-0.12).


CONCLUSIONS
Amniotic membrane promotes the proliferation on retinal Müller cells dose and time-dependently in vitro.
Hao
Yu-hua
YH
Department of Ophthalmology, Second Hospital of Hebei Medical University, Shijiazhuang 050000, China.
Ma
Jing-xue
JX
Xiu
He-ming
HM
Xu
Zheng
Z
D000650:Amnion; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005260:Female; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D046529:Microscopy, Electron, Transmission; D009457:Neuroglia / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D011817:Rabbits; D012160:Retina / Q000166:cytology; D009418:S100 Proteins / Q000032:analysis; D014020:Tissue Extracts / Q000494:pharmacology*
D005904:Glial Fibrillary Acidic Protein; D009418:S100 Proteins; D014020:Tissue Extracts