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Embryonic stem cell–derived photoreceptor precursor cells differentiated by coculture with RPE cells
PURPOSE: To describe the derivation of photoreceptor precursor cells from human embryonic stem cells by coculture with RPE cells. METHODS: Human embryonic stem cells were induced to differentiate into neural precursor cells and then cocultured with RPE cells to obtain cells showing retinal photorece...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Molecular Vision
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8116258/ https://www.ncbi.nlm.nih.gov/pubmed/34012231 |
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author | Shin, Joo Young Ma, DaeJoong Lim, Mi-Sun Cho, Myung Soo Kim, Young Joo Yu, Hyeong Gon |
author_facet | Shin, Joo Young Ma, DaeJoong Lim, Mi-Sun Cho, Myung Soo Kim, Young Joo Yu, Hyeong Gon |
author_sort | Shin, Joo Young |
collection | PubMed |
description | PURPOSE: To describe the derivation of photoreceptor precursor cells from human embryonic stem cells by coculture with RPE cells. METHODS: Human embryonic stem cells were induced to differentiate into neural precursor cells and then cocultured with RPE cells to obtain cells showing retinal photoreceptor features. Immunofluorescent staining, reverse transcription–PCR (RT–PCR), and microarray analysis were performed to identify photoreceptor markers, and a cGMP assay was used for in vitro functional analysis. After subretinal injection in rat animal models, retinal function was determined with electroretinography and optokinetic response detection, and immunofluorescent staining was performed to assess the survival of the injected cells. RESULTS: Cocultured cells were positive for rhodopsin, red and blue opsin, recoverin, and phosphodiesterase 6 beta on immunofluorescent staining and RT–PCR. Serial detection of stem cell–, neural precursor–, and photoreceptor-specific markers was noted in each stage of differentiation with microarray analysis. Increased cGMP hydrolysis in light-exposed conditions compared to that in dark conditions was observed. After the subretinal injection in the rats, preservation of optokinetic responses was noted up to 20 weeks, while electroretinographic response decreased. Survival of the injected cells was confirmed with positive immunofluorescence staining of human markers at 8 weeks. CONCLUSIONS: Cells showed photoreceptor-specific features when stem cell–derived neurogenic precursors were cocultured with RPE cells. |
format | Online Article Text |
id | pubmed-8116258 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Molecular Vision |
record_format | MEDLINE/PubMed |
spelling | pubmed-81162582021-05-18 Embryonic stem cell–derived photoreceptor precursor cells differentiated by coculture with RPE cells Shin, Joo Young Ma, DaeJoong Lim, Mi-Sun Cho, Myung Soo Kim, Young Joo Yu, Hyeong Gon Mol Vis Research Article PURPOSE: To describe the derivation of photoreceptor precursor cells from human embryonic stem cells by coculture with RPE cells. METHODS: Human embryonic stem cells were induced to differentiate into neural precursor cells and then cocultured with RPE cells to obtain cells showing retinal photoreceptor features. Immunofluorescent staining, reverse transcription–PCR (RT–PCR), and microarray analysis were performed to identify photoreceptor markers, and a cGMP assay was used for in vitro functional analysis. After subretinal injection in rat animal models, retinal function was determined with electroretinography and optokinetic response detection, and immunofluorescent staining was performed to assess the survival of the injected cells. RESULTS: Cocultured cells were positive for rhodopsin, red and blue opsin, recoverin, and phosphodiesterase 6 beta on immunofluorescent staining and RT–PCR. Serial detection of stem cell–, neural precursor–, and photoreceptor-specific markers was noted in each stage of differentiation with microarray analysis. Increased cGMP hydrolysis in light-exposed conditions compared to that in dark conditions was observed. After the subretinal injection in the rats, preservation of optokinetic responses was noted up to 20 weeks, while electroretinographic response decreased. Survival of the injected cells was confirmed with positive immunofluorescence staining of human markers at 8 weeks. CONCLUSIONS: Cells showed photoreceptor-specific features when stem cell–derived neurogenic precursors were cocultured with RPE cells. Molecular Vision 2021-05-09 /pmc/articles/PMC8116258/ /pubmed/34012231 Text en Copyright © 2021 Molecular Vision. https://creativecommons.org/licenses/by-nc-nd/3.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited, used for non-commercial purposes, and is not altered or transformed. |
spellingShingle | Research Article Shin, Joo Young Ma, DaeJoong Lim, Mi-Sun Cho, Myung Soo Kim, Young Joo Yu, Hyeong Gon Embryonic stem cell–derived photoreceptor precursor cells differentiated by coculture with RPE cells |
title | Embryonic stem cell–derived photoreceptor precursor cells differentiated by coculture with RPE cells |
title_full | Embryonic stem cell–derived photoreceptor precursor cells differentiated by coculture with RPE cells |
title_fullStr | Embryonic stem cell–derived photoreceptor precursor cells differentiated by coculture with RPE cells |
title_full_unstemmed | Embryonic stem cell–derived photoreceptor precursor cells differentiated by coculture with RPE cells |
title_short | Embryonic stem cell–derived photoreceptor precursor cells differentiated by coculture with RPE cells |
title_sort | embryonic stem cell–derived photoreceptor precursor cells differentiated by coculture with rpe cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8116258/ https://www.ncbi.nlm.nih.gov/pubmed/34012231 |
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