Effects of Y-27632, a Rho-associated Kinase Inhibitor, on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells

PURPOSE: Human corneal endothelial progenitor cells (HCEPs), which has been selectively isolated and differentiated into human corneal endothelial cells (HCECs), are crucial for repairing corneal endothelial damage. In this study, we evaluated the roles of a Rho-assisted kinase (ROCK) inhibitor, Y-2...

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Autores principales: Shin, Haeeun, Min, Joon Ki, Kim, Na Rae, Seo, Kyoung Yul, Chin, Hee Seung, Lee, Soyoung, Jung, Ji Won
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Korean Ophthalmological Society 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9935058/
https://www.ncbi.nlm.nih.gov/pubmed/36549303
http://dx.doi.org/10.3341/kjo.2022.0133
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author Shin, Haeeun
Min, Joon Ki
Kim, Na Rae
Seo, Kyoung Yul
Chin, Hee Seung
Lee, Soyoung
Jung, Ji Won
author_facet Shin, Haeeun
Min, Joon Ki
Kim, Na Rae
Seo, Kyoung Yul
Chin, Hee Seung
Lee, Soyoung
Jung, Ji Won
author_sort Shin, Haeeun
collection PubMed
description PURPOSE: Human corneal endothelial progenitor cells (HCEPs), which has been selectively isolated and differentiated into human corneal endothelial cells (HCECs), are crucial for repairing corneal endothelial damage. In this study, we evaluated the roles of a Rho-assisted kinase (ROCK) inhibitor, Y-27632, on the isolation and expansion of HCEPs, and assessed the in vitro effects of different concentrations of Y-27632 on the differentiated HCEPs. METHODS: HCEPs were isolated and expanded in a medium with and without 10μM Y-27632, and then differentiated into HCECs in a medium with fetal bovine serum. The characteristics of HCEPs and differentiated HCEPs were confirmed by immunofluorescence staining. The proliferation, viability, morphology, and wound-healing ability of differentiated HCEPs were assessed in the presence of different concentrations of Y-27632. RESULTS: Y-27632 enabled the isolation and expansion of HCEPs from the corneal endothelium. The differentiated HCEPs showed an optimal increase in proliferation and survival in the presence of 10μM Y-27632. As the concentration of Y-27632 increased, differentiated HCEPs became elongated, and actin filaments were redistributed to the periphery of cells. Y-27632 also caused a concentration-dependent enhancement in the wound-healing ability of differentiated HCEPs. CONCLUSIONS: Y-27632 enabled the isolation and expansion of HCEPs. It also enhanced the proliferation, viability, and migration of differentiated HCEPs.
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spelling pubmed-99350582023-02-17 Effects of Y-27632, a Rho-associated Kinase Inhibitor, on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells Shin, Haeeun Min, Joon Ki Kim, Na Rae Seo, Kyoung Yul Chin, Hee Seung Lee, Soyoung Jung, Ji Won Korean J Ophthalmol Original Article PURPOSE: Human corneal endothelial progenitor cells (HCEPs), which has been selectively isolated and differentiated into human corneal endothelial cells (HCECs), are crucial for repairing corneal endothelial damage. In this study, we evaluated the roles of a Rho-assisted kinase (ROCK) inhibitor, Y-27632, on the isolation and expansion of HCEPs, and assessed the in vitro effects of different concentrations of Y-27632 on the differentiated HCEPs. METHODS: HCEPs were isolated and expanded in a medium with and without 10μM Y-27632, and then differentiated into HCECs in a medium with fetal bovine serum. The characteristics of HCEPs and differentiated HCEPs were confirmed by immunofluorescence staining. The proliferation, viability, morphology, and wound-healing ability of differentiated HCEPs were assessed in the presence of different concentrations of Y-27632. RESULTS: Y-27632 enabled the isolation and expansion of HCEPs from the corneal endothelium. The differentiated HCEPs showed an optimal increase in proliferation and survival in the presence of 10μM Y-27632. As the concentration of Y-27632 increased, differentiated HCEPs became elongated, and actin filaments were redistributed to the periphery of cells. Y-27632 also caused a concentration-dependent enhancement in the wound-healing ability of differentiated HCEPs. CONCLUSIONS: Y-27632 enabled the isolation and expansion of HCEPs. It also enhanced the proliferation, viability, and migration of differentiated HCEPs. Korean Ophthalmological Society 2023-02 2022-12-22 /pmc/articles/PMC9935058/ /pubmed/36549303 http://dx.doi.org/10.3341/kjo.2022.0133 Text en © 2023 The Korean Ophthalmological Society https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access journal distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) ) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Shin, Haeeun
Min, Joon Ki
Kim, Na Rae
Seo, Kyoung Yul
Chin, Hee Seung
Lee, Soyoung
Jung, Ji Won
Effects of Y-27632, a Rho-associated Kinase Inhibitor, on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells
title Effects of Y-27632, a Rho-associated Kinase Inhibitor, on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells
title_full Effects of Y-27632, a Rho-associated Kinase Inhibitor, on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells
title_fullStr Effects of Y-27632, a Rho-associated Kinase Inhibitor, on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells
title_full_unstemmed Effects of Y-27632, a Rho-associated Kinase Inhibitor, on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells
title_short Effects of Y-27632, a Rho-associated Kinase Inhibitor, on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells
title_sort effects of y-27632, a rho-associated kinase inhibitor, on human corneal endothelial cells cultured by isolating human corneal endothelial progenitor cells
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9935058/
https://www.ncbi.nlm.nih.gov/pubmed/36549303
http://dx.doi.org/10.3341/kjo.2022.0133
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