RRC ID 51789
Author Wu MF, Stachon T, Langenbucher A, Seitz B, Szentmáry N.
Title Effect of Amniotic Membrane Suspension (AMS) and Amniotic Membrane Homogenate (AMH) on Human Corneal Epithelial Cell Viability, Migration and Proliferation In Vitro.
Journal Curr Eye Res
Abstract PURPOSE:To analyze the effects of different concentrations of amniotic membrane suspension (AMS) or amniotic membrane homogenate (AMH) on human corneal epithelial cell (HCEC) viability, migration and proliferation.
METHODS:Amniotic membranes (AMs) of 13 placentas were prepared and stored at -80°C. For AMS preparation, following de-freezing, AM pieces were inserted in six-well plates and 5 ml Dulbecco's Modified Eagle's Medium (DMEM)/F12 (with 5% fetal bovine serum, FBS) per gram tissue was added for 96 h. After removal of the AM, the remaining supernatant was collected for experiments. For AMH preparation, following de-freezing, AMs were homogenized in liquid nitrogen and 5 ml DMEM/F12 (with 5% FBS) per gram tissue was added. Following centrifugation, the supernatant was collected for experiments. HCECs were expanded and incubated in DMEM/F12, 5% FBS supplemented by 15%, 30% or 100% AMS or 15% or 30% AMH. Viability was analyzed using Cell Proliferation Kit XTT, migration using wound healing assay and proliferation by the cell proliferation ELISA BrdU kit.
RESULTS:HCEC viability remained unchanged using 15% or 30% AMS (p = 1.0 for both); however, it decreased significantly using 100% AMS (p < 0.001) or 15% (p = 0.041) or 30% AMH (p < 0.001), compared to controls. Using 15% or 30% AMS, HCEC migration increased significantly (p < 0.001 for both). Using 15% or 30% AMH (p = 0.153; p = 0.083), HCEC migration remained unchanged and 100% AMS inhibited HCEC migration (p < 0.001). Next, 15% and 30% AMS had no effect on HCEC proliferation (p = 0.454 and p = 0.119), but 100% AMS (p < 0.001) and 15% (p = 0.002) and 30% AMH (p = 0.001) inhibited HCEC proliferation significantly.
CONCLUSION:With unchanged HCEC viability and proliferation and increased HCEC migration, 15% and 30% AMS application seems to be the most appropriate method to support epithelial healing.
Volume 42(3)
Pages 351-357
Published 2017-3-1
DOI 10.1080/02713683.2016.1192193
PMID 27420558
MeSH Adult Amnion* Cell Movement / drug effects Cell Proliferation / drug effects Cells, Cultured Corneal Injuries / drug therapy* Corneal Injuries / metabolism Corneal Injuries / pathology Culture Media, Conditioned / pharmacology* Enzyme-Linked Immunosorbent Assay Epithelium, Corneal Female Humans Pregnancy Young Adult
IF 1.754
Times Cited 6
Resource
Human and Animal Cells HCE-T(RCB2280)