An NKX2-1 GFP and TP63 tdTomato dual fluorescent reporter for the investigation of human lung basal cell biology

Abstract Basal cells are multipotent stem cells responsible for the repair and regeneration of all the epithelial cell types present in the proximal lung. In mice, the elusive origins of basal cells and their contribution to lung development were recently revealed by high-resolution, lineage tracing...

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Autores principales:	Kim Jee Goh, Ee Kim Tan, Hao Lu, Sudipto Roy, N. Ray Dunn
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/469f529043274d93b41130ab02d7d2c7
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spelling	oai:doaj.org-article:469f529043274d93b41130ab02d7d2c72021-12-02T13:34:32ZAn NKX2-1 GFP and TP63 tdTomato dual fluorescent reporter for the investigation of human lung basal cell biology10.1038/s41598-021-83825-62045-2322https://doaj.org/article/469f529043274d93b41130ab02d7d2c72021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-83825-6https://doaj.org/toc/2045-2322Abstract Basal cells are multipotent stem cells responsible for the repair and regeneration of all the epithelial cell types present in the proximal lung. In mice, the elusive origins of basal cells and their contribution to lung development were recently revealed by high-resolution, lineage tracing studies. It however remains unclear if human basal cells originate and participate in lung development in a similar fashion, particularly with mounting evidence for significant species-specific differences in this process. To address this outstanding question, in the last several years differentiation protocols incorporating human pluripotent stem cells (hPSC) have been developed to produce human basal cells in vitro with varying efficiencies. To facilitate this endeavour, we introduced tdTomato into the human TP63 gene, whose expression specifically labels basal cells, in the background of a previously described hPSC line harbouring an NKX2-1 GFP reporter allele. The functionality and specificity of the NKX2-1 GFP ;TP63 tdTomato hPSC line was validated by directed differentiation into lung progenitors as well as more specialised lung epithelial subtypes using an organoid platform. This dual fluorescent reporter hPSC line will be useful for tracking, isolating and expanding basal cells from heterogenous differentiation cultures for further study.Kim Jee GohEe Kim TanHao LuSudipto RoyN. Ray DunnNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-17 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kim Jee Goh Ee Kim Tan Hao Lu Sudipto Roy N. Ray Dunn An NKX2-1 GFP and TP63 tdTomato dual fluorescent reporter for the investigation of human lung basal cell biology
description	Abstract Basal cells are multipotent stem cells responsible for the repair and regeneration of all the epithelial cell types present in the proximal lung. In mice, the elusive origins of basal cells and their contribution to lung development were recently revealed by high-resolution, lineage tracing studies. It however remains unclear if human basal cells originate and participate in lung development in a similar fashion, particularly with mounting evidence for significant species-specific differences in this process. To address this outstanding question, in the last several years differentiation protocols incorporating human pluripotent stem cells (hPSC) have been developed to produce human basal cells in vitro with varying efficiencies. To facilitate this endeavour, we introduced tdTomato into the human TP63 gene, whose expression specifically labels basal cells, in the background of a previously described hPSC line harbouring an NKX2-1 GFP reporter allele. The functionality and specificity of the NKX2-1 GFP ;TP63 tdTomato hPSC line was validated by directed differentiation into lung progenitors as well as more specialised lung epithelial subtypes using an organoid platform. This dual fluorescent reporter hPSC line will be useful for tracking, isolating and expanding basal cells from heterogenous differentiation cultures for further study.
format	article
author	Kim Jee Goh Ee Kim Tan Hao Lu Sudipto Roy N. Ray Dunn
author_facet	Kim Jee Goh Ee Kim Tan Hao Lu Sudipto Roy N. Ray Dunn
author_sort	Kim Jee Goh
title	An NKX2-1 GFP and TP63 tdTomato dual fluorescent reporter for the investigation of human lung basal cell biology
title_short	An NKX2-1 GFP and TP63 tdTomato dual fluorescent reporter for the investigation of human lung basal cell biology
title_full	An NKX2-1 GFP and TP63 tdTomato dual fluorescent reporter for the investigation of human lung basal cell biology
title_fullStr	An NKX2-1 GFP and TP63 tdTomato dual fluorescent reporter for the investigation of human lung basal cell biology
title_full_unstemmed	An NKX2-1 GFP and TP63 tdTomato dual fluorescent reporter for the investigation of human lung basal cell biology
title_sort	nkx2-1 gfp and tp63 tdtomato dual fluorescent reporter for the investigation of human lung basal cell biology
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/469f529043274d93b41130ab02d7d2c7
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An NKX2-1 GFP and TP63 tdTomato dual fluorescent reporter for the investigation of human lung basal cell biology

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