論文 - 詳細
RRC ID | 51789 |
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著者 | Wu MF, Stachon T, Langenbucher A, Seitz B, Szentmáry N. |
タイトル | Effect of Amniotic Membrane Suspension (AMS) and Amniotic Membrane Homogenate (AMH) on Human Corneal Epithelial Cell Viability, Migration and Proliferation In Vitro. |
ジャーナル | 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. |
巻・号 | 42(3) |
ページ | 351-357 |
公開日 | 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 |
引用数 | 6 |
リソース情報 | |
ヒト・動物細胞 | HCE-T(RCB2280) |