An in vivo gene delivery approach for the isolation of reasonable numbers of type 2 innate lymphoid cells
Group 2 innate lymphoid cells (ILC2s) are a recently recognized subset of innate lymphocytes with crucial role in mucosal immunity and tissue homeostasis. Over the past decade, substantial advances in our understanding of ILC2 biology have established them as an essential element in innate and adapt...
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doaj-fbb0f95a59e1438d81e059714c7457b12021-01-02T05:10:55ZengElsevierMethodsX2215-01612020-01-017101054An in vivo gene delivery approach for the isolation of reasonable numbers of type 2 innate lymphoid cellsMichael Frech0Lisa Knipfer1Stefan Wirtz2Mario M. Zaiss3Department of Internal Medicine 3, Rheumatology and Immunology, Friedrich-Alexander-University Erlangen-Nürnberg (FAU) and Universitätsklinikum Erlangen, Erlangen, Germany; Deutsches Zentrum für Immuntherapie (DZI), GermanyDepartment of Internal Medicine 1, University of Erlangen-Nuremberg, Germany; Deutsches Zentrum für Immuntherapie (DZI), GermanyDepartment of Internal Medicine 1, University of Erlangen-Nuremberg, Germany; Deutsches Zentrum für Immuntherapie (DZI), GermanyDepartment of Internal Medicine 3, Rheumatology and Immunology, Friedrich-Alexander-University Erlangen-Nürnberg (FAU) and Universitätsklinikum Erlangen, Erlangen, Germany; Department of Internal Medicine 1, University of Erlangen-Nuremberg, Germany; Corresponding author.Group 2 innate lymphoid cells (ILC2s) are a recently recognized subset of innate lymphocytes with crucial role in mucosal immunity and tissue homeostasis. Over the past decade, substantial advances in our understanding of ILC2 biology have established them as an essential element in innate and adaptive immunity. However, their relatively low abundance and laborious purification from mucosal tissues make their study difficult. Moreover, due to a lack of an ILC2-specific Cre mouse-line, adoptive transfer of ILC2s into ILC-deficient hosts is inevitable. Herein we describe an in-depth protocol for the induction, isolation, and expansion of murine ILC2s. By combining an in vivo gene delivery approach to boost ILC2 numbers and a cell culture strategy to expand isolated cells, large quantities of highly pure ILC2s can be obtained. The isolated cells maintain their phenotype and can be used for subsequent cell transfer or in vitro studies. In comparison to previous protocols, this approach is cost-effective and efficient with potential yield of more than 20 million ILC2s isolated per mouse.• Group 2 innate lymphoid cells (ILC2s) are extensively studied in mouse models and humans in recent years.• Low abundance of ILC2s and current lack of specific ILC2 knockout mice makes in vivo research challenging.• This method allows high and pure ILC2 numbers for in vitro or adoptive in vivo transfer experiments.http://www.sciencedirect.com/science/article/pii/S2215016120302740Isolation and expansion of murine ILC2s |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Michael Frech Lisa Knipfer Stefan Wirtz Mario M. Zaiss |
spellingShingle |
Michael Frech Lisa Knipfer Stefan Wirtz Mario M. Zaiss An in vivo gene delivery approach for the isolation of reasonable numbers of type 2 innate lymphoid cells MethodsX Isolation and expansion of murine ILC2s |
author_facet |
Michael Frech Lisa Knipfer Stefan Wirtz Mario M. Zaiss |
author_sort |
Michael Frech |
title |
An in vivo gene delivery approach for the isolation of reasonable numbers of type 2 innate lymphoid cells |
title_short |
An in vivo gene delivery approach for the isolation of reasonable numbers of type 2 innate lymphoid cells |
title_full |
An in vivo gene delivery approach for the isolation of reasonable numbers of type 2 innate lymphoid cells |
title_fullStr |
An in vivo gene delivery approach for the isolation of reasonable numbers of type 2 innate lymphoid cells |
title_full_unstemmed |
An in vivo gene delivery approach for the isolation of reasonable numbers of type 2 innate lymphoid cells |
title_sort |
in vivo gene delivery approach for the isolation of reasonable numbers of type 2 innate lymphoid cells |
publisher |
Elsevier |
series |
MethodsX |
issn |
2215-0161 |
publishDate |
2020-01-01 |
description |
Group 2 innate lymphoid cells (ILC2s) are a recently recognized subset of innate lymphocytes with crucial role in mucosal immunity and tissue homeostasis. Over the past decade, substantial advances in our understanding of ILC2 biology have established them as an essential element in innate and adaptive immunity. However, their relatively low abundance and laborious purification from mucosal tissues make their study difficult. Moreover, due to a lack of an ILC2-specific Cre mouse-line, adoptive transfer of ILC2s into ILC-deficient hosts is inevitable. Herein we describe an in-depth protocol for the induction, isolation, and expansion of murine ILC2s. By combining an in vivo gene delivery approach to boost ILC2 numbers and a cell culture strategy to expand isolated cells, large quantities of highly pure ILC2s can be obtained. The isolated cells maintain their phenotype and can be used for subsequent cell transfer or in vitro studies. In comparison to previous protocols, this approach is cost-effective and efficient with potential yield of more than 20 million ILC2s isolated per mouse.• Group 2 innate lymphoid cells (ILC2s) are extensively studied in mouse models and humans in recent years.• Low abundance of ILC2s and current lack of specific ILC2 knockout mice makes in vivo research challenging.• This method allows high and pure ILC2 numbers for in vitro or adoptive in vivo transfer experiments. |
topic |
Isolation and expansion of murine ILC2s |
url |
http://www.sciencedirect.com/science/article/pii/S2215016120302740 |
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