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In vitro differentiation of cGMP-grade retinal pigmented epithelium from human embryonic stem cells
BACKGROUND: The World Health Organization (WHO) estimates that the number of individuals who lose their vision due to retinal degeneration is expected to reach 6 million annually in 2020. The retinal degenerative diseases affect the macula, which is responsible for central and detailed vision. Most...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6802162/ https://www.ncbi.nlm.nih.gov/pubmed/31646003 http://dx.doi.org/10.1186/s40942-019-0194-7 |
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author | Lojudice, Fernando H. Fernandes, Rodrigo A. Brant Innocenti, Francesco Franciozi, Carlos E. Cristovam, Priscila Maia, Maurício Sogayar, Mari C. Belfort, Rubens |
author_facet | Lojudice, Fernando H. Fernandes, Rodrigo A. Brant Innocenti, Francesco Franciozi, Carlos E. Cristovam, Priscila Maia, Maurício Sogayar, Mari C. Belfort, Rubens |
author_sort | Lojudice, Fernando H. |
collection | PubMed |
description | BACKGROUND: The World Health Organization (WHO) estimates that the number of individuals who lose their vision due to retinal degeneration is expected to reach 6 million annually in 2020. The retinal degenerative diseases affect the macula, which is responsible for central and detailed vision. Most macular degeneration, i.e., age-related macular degeneration (AMD) develops in the elderly; however, certain hereditary diseases, such as the Stargardt disease, also affect young people. This degeneration begins with loss of retinal pigmented epithelium (RPE) due to formation of drusen (atrophic) or abnormal vessels (exudative). In wet AMD, numerous drugs are available to successful treat the disease; however, no proven therapy currently is available to treat dry AMD or Stargardt. Since its discovery, human embryonic stem cells (hESCs) have been considered a valuable therapeutic tool. Some evidence has shown that transplantation of RPEs differentiated from hESCs cells can result in recovery of both RPE and photoreceptors and prevent visual loss. METHODS: The human embryonic WA-09 stem cell lineage was cultured under current Good Manufacturing Practices (cGMP) conditions using serum-free media and supplements. The colonies were isolated manually and allowed to spontaneously differentiate into RPE cells. RESULTS: This simple and effective protocol required minimal manipulation and yielded more than 10e8 RPE cells by the end of the differentiation and enrichment processes, with cells exhibiting a cobblestone morphology and displaying cellular markers and a gene expression profile typical of mature RPE cells. Moreover, the differentiated cells displayed phagocytic activity and only a small percentage of the total cells remained positive for the Octamer-binding transcriptions factor 4 (OCT-4) pluripotency cell marker. CONCLUSIONS: These results showed that functional RPE cells can be produced efficiently and suggested the possibility of scaling-up to aim at therapeutic protocols for retinal diseases associated with RPE degeneration. |
format | Online Article Text |
id | pubmed-6802162 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-68021622019-10-23 In vitro differentiation of cGMP-grade retinal pigmented epithelium from human embryonic stem cells Lojudice, Fernando H. Fernandes, Rodrigo A. Brant Innocenti, Francesco Franciozi, Carlos E. Cristovam, Priscila Maia, Maurício Sogayar, Mari C. Belfort, Rubens Int J Retina Vitreous Original Article BACKGROUND: The World Health Organization (WHO) estimates that the number of individuals who lose their vision due to retinal degeneration is expected to reach 6 million annually in 2020. The retinal degenerative diseases affect the macula, which is responsible for central and detailed vision. Most macular degeneration, i.e., age-related macular degeneration (AMD) develops in the elderly; however, certain hereditary diseases, such as the Stargardt disease, also affect young people. This degeneration begins with loss of retinal pigmented epithelium (RPE) due to formation of drusen (atrophic) or abnormal vessels (exudative). In wet AMD, numerous drugs are available to successful treat the disease; however, no proven therapy currently is available to treat dry AMD or Stargardt. Since its discovery, human embryonic stem cells (hESCs) have been considered a valuable therapeutic tool. Some evidence has shown that transplantation of RPEs differentiated from hESCs cells can result in recovery of both RPE and photoreceptors and prevent visual loss. METHODS: The human embryonic WA-09 stem cell lineage was cultured under current Good Manufacturing Practices (cGMP) conditions using serum-free media and supplements. The colonies were isolated manually and allowed to spontaneously differentiate into RPE cells. RESULTS: This simple and effective protocol required minimal manipulation and yielded more than 10e8 RPE cells by the end of the differentiation and enrichment processes, with cells exhibiting a cobblestone morphology and displaying cellular markers and a gene expression profile typical of mature RPE cells. Moreover, the differentiated cells displayed phagocytic activity and only a small percentage of the total cells remained positive for the Octamer-binding transcriptions factor 4 (OCT-4) pluripotency cell marker. CONCLUSIONS: These results showed that functional RPE cells can be produced efficiently and suggested the possibility of scaling-up to aim at therapeutic protocols for retinal diseases associated with RPE degeneration. BioMed Central 2019-10-21 /pmc/articles/PMC6802162/ /pubmed/31646003 http://dx.doi.org/10.1186/s40942-019-0194-7 Text en © The Author(s) 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Original Article Lojudice, Fernando H. Fernandes, Rodrigo A. Brant Innocenti, Francesco Franciozi, Carlos E. Cristovam, Priscila Maia, Maurício Sogayar, Mari C. Belfort, Rubens In vitro differentiation of cGMP-grade retinal pigmented epithelium from human embryonic stem cells |
title | In vitro differentiation of cGMP-grade retinal pigmented epithelium from human embryonic stem cells |
title_full | In vitro differentiation of cGMP-grade retinal pigmented epithelium from human embryonic stem cells |
title_fullStr | In vitro differentiation of cGMP-grade retinal pigmented epithelium from human embryonic stem cells |
title_full_unstemmed | In vitro differentiation of cGMP-grade retinal pigmented epithelium from human embryonic stem cells |
title_short | In vitro differentiation of cGMP-grade retinal pigmented epithelium from human embryonic stem cells |
title_sort | in vitro differentiation of cgmp-grade retinal pigmented epithelium from human embryonic stem cells |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6802162/ https://www.ncbi.nlm.nih.gov/pubmed/31646003 http://dx.doi.org/10.1186/s40942-019-0194-7 |
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