Serum free culture for the expansion and study of type 2 innate lymphoid cells

Abstract Type 2 innate lymphoid cells (ILC2s) were discovered approximately ten years ago and their clinical relevance is gaining greater importance. However, their successful isolation from mammalian tissues and in vitro culture and expansion continues to pose challenges. This is partly due to thei...

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Autores principales:	Pablo de Lucía Finkel, Christopher Sherwood, Iryna Saranchova, Wenjing Xia, Lonna Munro, Cheryl G. Pfeifer, James M. Piret, Wilfred A. Jefferies
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/96f8a16afbfc41198379d0874751a533
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spelling	oai:doaj.org-article:96f8a16afbfc41198379d0874751a5332021-12-02T17:47:37ZSerum free culture for the expansion and study of type 2 innate lymphoid cells10.1038/s41598-021-91500-z2045-2322https://doaj.org/article/96f8a16afbfc41198379d0874751a5332021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91500-zhttps://doaj.org/toc/2045-2322Abstract Type 2 innate lymphoid cells (ILC2s) were discovered approximately ten years ago and their clinical relevance is gaining greater importance. However, their successful isolation from mammalian tissues and in vitro culture and expansion continues to pose challenges. This is partly due to their scarcity compared to other leukocyte populations, but also because our current knowledge of ILC2 biology is incomplete. This study is focused on ST2+ IL-25Rlo lung resident ILC2s and demonstrate for the first time a methodology allowing mouse type 2 innate lymphoid cells to be cultured, and their numbers expanded in serum-free medium supplemented with Interleukins IL-33, IL-2, IL-7 and TSLP. The procedures described methods to isolate ILC2s and support their growth for up to a week while maintaining their phenotype. During this time, they significantly expand from low to high cell concentrations. Furthermore, for the first time, sub-cultures of primary ILC2 purifications in larger 24- and 6-well plates were undertaken in order to compare their growth in other media. In culture, ILC2s had doubling times of 21 h, a growth rate of 0.032 h−1 and could be sub-cultured in early or late phases of exponential growth. These studies form the basis for expanding ILC2 populations that will facilitate the study and potential applications of these rare cells under defined, serum-free conditions.Pablo de Lucía FinkelChristopher SherwoodIryna SaranchovaWenjing XiaLonna MunroCheryl G. PfeiferJames M. PiretWilfred A. JefferiesNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Pablo de Lucía Finkel Christopher Sherwood Iryna Saranchova Wenjing Xia Lonna Munro Cheryl G. Pfeifer James M. Piret Wilfred A. Jefferies Serum free culture for the expansion and study of type 2 innate lymphoid cells
description	Abstract Type 2 innate lymphoid cells (ILC2s) were discovered approximately ten years ago and their clinical relevance is gaining greater importance. However, their successful isolation from mammalian tissues and in vitro culture and expansion continues to pose challenges. This is partly due to their scarcity compared to other leukocyte populations, but also because our current knowledge of ILC2 biology is incomplete. This study is focused on ST2+ IL-25Rlo lung resident ILC2s and demonstrate for the first time a methodology allowing mouse type 2 innate lymphoid cells to be cultured, and their numbers expanded in serum-free medium supplemented with Interleukins IL-33, IL-2, IL-7 and TSLP. The procedures described methods to isolate ILC2s and support their growth for up to a week while maintaining their phenotype. During this time, they significantly expand from low to high cell concentrations. Furthermore, for the first time, sub-cultures of primary ILC2 purifications in larger 24- and 6-well plates were undertaken in order to compare their growth in other media. In culture, ILC2s had doubling times of 21 h, a growth rate of 0.032 h−1 and could be sub-cultured in early or late phases of exponential growth. These studies form the basis for expanding ILC2 populations that will facilitate the study and potential applications of these rare cells under defined, serum-free conditions.
format	article
author	Pablo de Lucía Finkel Christopher Sherwood Iryna Saranchova Wenjing Xia Lonna Munro Cheryl G. Pfeifer James M. Piret Wilfred A. Jefferies
author_facet	Pablo de Lucía Finkel Christopher Sherwood Iryna Saranchova Wenjing Xia Lonna Munro Cheryl G. Pfeifer James M. Piret Wilfred A. Jefferies
author_sort	Pablo de Lucía Finkel
title	Serum free culture for the expansion and study of type 2 innate lymphoid cells
title_short	Serum free culture for the expansion and study of type 2 innate lymphoid cells
title_full	Serum free culture for the expansion and study of type 2 innate lymphoid cells
title_fullStr	Serum free culture for the expansion and study of type 2 innate lymphoid cells
title_full_unstemmed	Serum free culture for the expansion and study of type 2 innate lymphoid cells
title_sort	serum free culture for the expansion and study of type 2 innate lymphoid cells
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/96f8a16afbfc41198379d0874751a533
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Serum free culture for the expansion and study of type 2 innate lymphoid cells

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