Transcriptomic Profiling of Human Pluripotent Stem Cell-derived Retinal Pigment Epithelium over Time

Human pluripotent stem cell (hPSC)-derived progenies are immature versions of cells, presenting a potential limitation to the accurate modelling of diseases associated with maturity or age. Hence, it is important to characterise how closely cells used in culture resemble their native counterparts. I...

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Autores principales:	Grace E. Lidgerwood, Anne Senabouth, Casey J.A. Smith-Anttila, Vikkitharan Gnanasambandapillai, Dominik C. Kaczorowski, Daniela Amann-Zalcenstein, Erica L. Fletcher, Shalin H. Naik, Alex W. Hewitt, Joseph E. Powell, Alice Pébay
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Human embryonic stem cell Human pluripotent stem cell Retinal pigment epithelium Single-cell RNA sequencing Ageing Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/a1b752ada11e4c119fcf167e7656b297
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spelling	oai:doaj.org-article:a1b752ada11e4c119fcf167e7656b2972021-11-16T04:09:19ZTranscriptomic Profiling of Human Pluripotent Stem Cell-derived Retinal Pigment Epithelium over Time1672-022910.1016/j.gpb.2020.08.002https://doaj.org/article/a1b752ada11e4c119fcf167e7656b2972021-04-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S1672022920301352https://doaj.org/toc/1672-0229Human pluripotent stem cell (hPSC)-derived progenies are immature versions of cells, presenting a potential limitation to the accurate modelling of diseases associated with maturity or age. Hence, it is important to characterise how closely cells used in culture resemble their native counterparts. In order to select appropriate time points of retinal pigment epithelium (RPE) cultures that reflect native counterparts, we characterised the transcriptomic profiles of the hPSC-derived RPE cells from 1- and 12-month cultures. We differentiated the human embryonic stem cell line H9 into RPE cells, performed single-cell RNA-sequencing of a total of 16,576 cells to assess the molecular changes of the RPE cells across these two culture time points. Our results indicate the stability of the RPE transcriptomic signature, with no evidence of an epithelial–mesenchymal transition, and with the maturing populations of the RPE observed with time in culture. Assessment of Gene Ontology pathways revealed that as the cultures age, RPE cells upregulate expression of genes involved in metal binding and antioxidant functions. This might reflect an increased ability to handle oxidative stress as cells mature. Comparison with native human RPE data confirms a maturing transcriptional profile of RPE cells in culture. These results suggest that long-term in vitro culture of RPE cells allows the modelling of specific phenotypes observed in native mature tissues. Our work highlights the transcriptional landscape of hPSC-derived RPE cells as they age in culture, which provides a reference for native and patient samples to be benchmarked against.Grace E. LidgerwoodAnne SenabouthCasey J.A. Smith-AnttilaVikkitharan GnanasambandapillaiDominik C. KaczorowskiDaniela Amann-ZalcensteinErica L. FletcherShalin H. NaikAlex W. HewittJoseph E. PowellAlice PébayElsevierarticleHuman embryonic stem cellHuman pluripotent stem cellRetinal pigment epitheliumSingle-cell RNA sequencingAgeingBiology (General)QH301-705.5ENGenomics, Proteomics & Bioinformatics, Vol 19, Iss 2, Pp 223-242 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Human embryonic stem cell Human pluripotent stem cell Retinal pigment epithelium Single-cell RNA sequencing Ageing Biology (General) QH301-705.5
spellingShingle	Human embryonic stem cell Human pluripotent stem cell Retinal pigment epithelium Single-cell RNA sequencing Ageing Biology (General) QH301-705.5 Grace E. Lidgerwood Anne Senabouth Casey J.A. Smith-Anttila Vikkitharan Gnanasambandapillai Dominik C. Kaczorowski Daniela Amann-Zalcenstein Erica L. Fletcher Shalin H. Naik Alex W. Hewitt Joseph E. Powell Alice Pébay Transcriptomic Profiling of Human Pluripotent Stem Cell-derived Retinal Pigment Epithelium over Time
description	Human pluripotent stem cell (hPSC)-derived progenies are immature versions of cells, presenting a potential limitation to the accurate modelling of diseases associated with maturity or age. Hence, it is important to characterise how closely cells used in culture resemble their native counterparts. In order to select appropriate time points of retinal pigment epithelium (RPE) cultures that reflect native counterparts, we characterised the transcriptomic profiles of the hPSC-derived RPE cells from 1- and 12-month cultures. We differentiated the human embryonic stem cell line H9 into RPE cells, performed single-cell RNA-sequencing of a total of 16,576 cells to assess the molecular changes of the RPE cells across these two culture time points. Our results indicate the stability of the RPE transcriptomic signature, with no evidence of an epithelial–mesenchymal transition, and with the maturing populations of the RPE observed with time in culture. Assessment of Gene Ontology pathways revealed that as the cultures age, RPE cells upregulate expression of genes involved in metal binding and antioxidant functions. This might reflect an increased ability to handle oxidative stress as cells mature. Comparison with native human RPE data confirms a maturing transcriptional profile of RPE cells in culture. These results suggest that long-term in vitro culture of RPE cells allows the modelling of specific phenotypes observed in native mature tissues. Our work highlights the transcriptional landscape of hPSC-derived RPE cells as they age in culture, which provides a reference for native and patient samples to be benchmarked against.
format	article
author	Grace E. Lidgerwood Anne Senabouth Casey J.A. Smith-Anttila Vikkitharan Gnanasambandapillai Dominik C. Kaczorowski Daniela Amann-Zalcenstein Erica L. Fletcher Shalin H. Naik Alex W. Hewitt Joseph E. Powell Alice Pébay
author_facet	Grace E. Lidgerwood Anne Senabouth Casey J.A. Smith-Anttila Vikkitharan Gnanasambandapillai Dominik C. Kaczorowski Daniela Amann-Zalcenstein Erica L. Fletcher Shalin H. Naik Alex W. Hewitt Joseph E. Powell Alice Pébay
author_sort	Grace E. Lidgerwood
title	Transcriptomic Profiling of Human Pluripotent Stem Cell-derived Retinal Pigment Epithelium over Time
title_short	Transcriptomic Profiling of Human Pluripotent Stem Cell-derived Retinal Pigment Epithelium over Time
title_full	Transcriptomic Profiling of Human Pluripotent Stem Cell-derived Retinal Pigment Epithelium over Time
title_fullStr	Transcriptomic Profiling of Human Pluripotent Stem Cell-derived Retinal Pigment Epithelium over Time
title_full_unstemmed	Transcriptomic Profiling of Human Pluripotent Stem Cell-derived Retinal Pigment Epithelium over Time
title_sort	transcriptomic profiling of human pluripotent stem cell-derived retinal pigment epithelium over time
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/a1b752ada11e4c119fcf167e7656b297
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Transcriptomic Profiling of Human Pluripotent Stem Cell-derived Retinal Pigment Epithelium over Time

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