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Small-Molecule Induction Promotes Corneal Endothelial Cell Differentiation From Human iPS Cells

Purpose: Corneal endothelial cells (CECs) serve as a barrier and foothold for the corneal stroma to maintain the function and transparency of the cornea. Loss of CECs during aging or disease states leads to blindness, and cell replacement therapy using either donated or artificially differentiated C...

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Autores principales: Chen, Jie, Ou, Qingjian, Wang, Zhe, Liu, Yifan, Hu, Shuqin, Liu, Yumeilan, Tian, Haibin, Xu, Jingying, Gao, Furong, Lu, Lixia, Jin, Caixia, Xu, Guo-Tong, Cui, Hong-Ping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8714889/
https://www.ncbi.nlm.nih.gov/pubmed/34976977
http://dx.doi.org/10.3389/fbioe.2021.788987
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author Chen, Jie
Ou, Qingjian
Wang, Zhe
Liu, Yifan
Hu, Shuqin
Liu, Yumeilan
Tian, Haibin
Xu, Jingying
Gao, Furong
Lu, Lixia
Jin, Caixia
Xu, Guo-Tong
Cui, Hong-Ping
author_facet Chen, Jie
Ou, Qingjian
Wang, Zhe
Liu, Yifan
Hu, Shuqin
Liu, Yumeilan
Tian, Haibin
Xu, Jingying
Gao, Furong
Lu, Lixia
Jin, Caixia
Xu, Guo-Tong
Cui, Hong-Ping
author_sort Chen, Jie
collection PubMed
description Purpose: Corneal endothelial cells (CECs) serve as a barrier and foothold for the corneal stroma to maintain the function and transparency of the cornea. Loss of CECs during aging or disease states leads to blindness, and cell replacement therapy using either donated or artificially differentiated CECs remains the only curative approach. Methods: Human induced pluripotent stem cells (hiPSCs) that were cultured in chemically defined medium were induced with dual-SMAD inhibition to differentiate into neural crest cells (NCCs). A small-molecule library was screened to differentiate the NCCs into corneal endothelial-like cells. The characteristics of these cells were identified with real-time PCR and immunofluorescence. Western blotting was applied to detect the signaling pathways and key factors regulated by the small molecules. Results: We developed an effective protocol to differentiate hiPSCs into CECs with defined small molecules. The hiPSC-CECs were characterized by ZO-1, AQP1, Vimentin and Na(+)/K(+)-ATPase. Based on our small-molecule screen, we identified a small-molecule combination, A769662 and AT13148, that enabled the most efficient production of CECs. The combination of A769662 and AT13148 upregulated the PKA/AKT signaling pathway, FOXO1 and PITX2 to promote the conversion of NCCs to CECs. Conclusion: We established an efficient small molecule-based method to differentiate hiPSCs into corneal endothelial-like cells, which might facilitate drug discovery and the development of cell-based therapies for corneal diseases.
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spelling pubmed-87148892021-12-30 Small-Molecule Induction Promotes Corneal Endothelial Cell Differentiation From Human iPS Cells Chen, Jie Ou, Qingjian Wang, Zhe Liu, Yifan Hu, Shuqin Liu, Yumeilan Tian, Haibin Xu, Jingying Gao, Furong Lu, Lixia Jin, Caixia Xu, Guo-Tong Cui, Hong-Ping Front Bioeng Biotechnol Bioengineering and Biotechnology Purpose: Corneal endothelial cells (CECs) serve as a barrier and foothold for the corneal stroma to maintain the function and transparency of the cornea. Loss of CECs during aging or disease states leads to blindness, and cell replacement therapy using either donated or artificially differentiated CECs remains the only curative approach. Methods: Human induced pluripotent stem cells (hiPSCs) that were cultured in chemically defined medium were induced with dual-SMAD inhibition to differentiate into neural crest cells (NCCs). A small-molecule library was screened to differentiate the NCCs into corneal endothelial-like cells. The characteristics of these cells were identified with real-time PCR and immunofluorescence. Western blotting was applied to detect the signaling pathways and key factors regulated by the small molecules. Results: We developed an effective protocol to differentiate hiPSCs into CECs with defined small molecules. The hiPSC-CECs were characterized by ZO-1, AQP1, Vimentin and Na(+)/K(+)-ATPase. Based on our small-molecule screen, we identified a small-molecule combination, A769662 and AT13148, that enabled the most efficient production of CECs. The combination of A769662 and AT13148 upregulated the PKA/AKT signaling pathway, FOXO1 and PITX2 to promote the conversion of NCCs to CECs. Conclusion: We established an efficient small molecule-based method to differentiate hiPSCs into corneal endothelial-like cells, which might facilitate drug discovery and the development of cell-based therapies for corneal diseases. Frontiers Media S.A. 2021-12-15 /pmc/articles/PMC8714889/ /pubmed/34976977 http://dx.doi.org/10.3389/fbioe.2021.788987 Text en Copyright © 2021 Chen, Ou, Wang, Liu, Hu, Liu, Tian, Xu, Gao, Lu, Jin, Xu and Cui. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Chen, Jie
Ou, Qingjian
Wang, Zhe
Liu, Yifan
Hu, Shuqin
Liu, Yumeilan
Tian, Haibin
Xu, Jingying
Gao, Furong
Lu, Lixia
Jin, Caixia
Xu, Guo-Tong
Cui, Hong-Ping
Small-Molecule Induction Promotes Corneal Endothelial Cell Differentiation From Human iPS Cells
title Small-Molecule Induction Promotes Corneal Endothelial Cell Differentiation From Human iPS Cells
title_full Small-Molecule Induction Promotes Corneal Endothelial Cell Differentiation From Human iPS Cells
title_fullStr Small-Molecule Induction Promotes Corneal Endothelial Cell Differentiation From Human iPS Cells
title_full_unstemmed Small-Molecule Induction Promotes Corneal Endothelial Cell Differentiation From Human iPS Cells
title_short Small-Molecule Induction Promotes Corneal Endothelial Cell Differentiation From Human iPS Cells
title_sort small-molecule induction promotes corneal endothelial cell differentiation from human ips cells
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8714889/
https://www.ncbi.nlm.nih.gov/pubmed/34976977
http://dx.doi.org/10.3389/fbioe.2021.788987
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