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Isolation, Cryopreservation and Culture of Human Amnion Epithelial Cells for Clinical Applications
Published on: December 21, 2014
A study on the preservation of fresh amniotic membrane
1Zhongshan Ophthalmic Center, Sun Yat-Sen University of Medical Sciences, Guangzhou 510060, China.
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 degrees C or cultured in DMEM at 37 degrees C, 5% CO2. 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 degrees C and 1 week in tissue culture for clinical use.
