A Study on the Preservation of Fresh Amniotic Membrane

Objective: To establish the standard preservation methods of fresh amniotic membranefor clinical use.Methods: Human placentas were collected aseptically from selective caesarean sectionsin normal women in time. Amniotic or placental membrane were peeled and preserved inN.S, P.B. SorDMEM at4°C or cultured in DMEM at 37°C, 5% CO2. Trypan-bluestaining, light and electronic microscopy were observed every six hours after preservation.Results: Seventy percent of amniotic epithelial cells survived after preservation in N. Sfor 6 hours, PBS 12 hours, DMEM 24 hours and 1 week in tissue culture. The amountof living epithelial cells maintained in placental membrane preservation was less thanthat in amniotic membrane preservation at the same time (t-test, P ＜ 0. 01) . Nocollagen degeneration was found during preservation.Conclusion: Preservative solution and time will affect the maintenance time of freshamniotic membrane greatly. Fresh amniotic membrane should be preserved within 6hours in N.S, 12 hours in P.B.S, 24 hours in DMEM at 4 °C and 1 week in tissteculture for clinical use.

Objective: To establish the standard preservation methods of fresh amniotic membrane for clinical use.Methods: Human placentas were collected aseptically from selective caesarean sections in normal women in time. Amniotic or placental membrane were peeled and preserved in N. S, P. B. S or DMEM at 4℃ or cultured in DMEM at 37℃, 5 % C02. Trypan-blue staining, light and electronic microscopy were observed every six hours after preservation. Results: Seventy percent of amniotic epithelial cells survived after preservation in N. S for 6 hours, PBS 12 hours, DMEM 24 hours and 1 week in tissue culture. The amount of living epithelial cells maintained in placental membrane preservation was less than that in amniotic membrane preservation at the same time (t-test, P <0.01) . No collagen degeneration was found during preservation.Conclusion: Preservative solution and time will affect the maintenance time of fresh amniotic membrane greatly. Fresh amniotic membrane should be preserved within 6 hours in N. S, 12 hours in P. B. S, 24 hours in DMEM at 4℃ and 1 week in tissue culture for clinical use. Eye Science 2001; 17 ; 158 ~ 162.