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Isolation, Cryopreservation and Culture of Human Amnion Epithelial Cells for Clinical Applications
Published on: December 21, 2014
[The effect of frozen storage for amniotic membrane ultrastructure]
1Ophthalmology Department, Ruijin Hospital Affiliated Shanghai Second Medical University, Shanghai 200025, China.
Objective:
To observe the effect of frozen storage for amniotic membrane ultrastructure, and provide the experimental evidence for clinical use.
Methods:
The amniotic membranes of fresh-obtained and -80 degrees C frozen storage 30 d, 60 d, 90 d and 180 d were examined with transmission electron microscope, and the ultrastructural changes were observed.
Results:
The epithelium in the fresh-obtained amniotic membrane was observed as intact structure, abundant organelles in cytoplasm and intercellular contact with desmosome. A great quantity of collagen micro-fibrils were found in the stroma, and the light and shade striation were observed in the collagen micro-fibrils of fresh-obtained amniotic membrane. The epithelium degenerated and died gradually as the increase of frozen storage time. At the frozen storage of 90 days, the epithelial chromatin dissolved, organelles degenerated and mitochondrion vacuolated. At the frozen storage of 30 days and 60 days, the pattern of collagen micro-fibrils in stoma was similar to that in fresh-obtained amniotic membrane; At the frozen storage of 90 days, some of collagen micro-fibrils in stoma showed intumescence. At the frozen storage of 180 days, the number of collagen micro-fibrils in stoma decreased, and some of them showed intumescence and dissolvent.
Conclusion:
Frozen storage can result in amniotic membrane epithelium death and collagen micro-fibrils intumescence and dissolvent.
