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NADPH Oxidases pp 587-622 | Cite as

Ex Vivo Models of Chronic Granulomatous Disease

  • Julie Brault
  • Bénédicte Vigne
  • Marie José StasiaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1982)

Abstract

Induced pluripotent stem cells (iPSCs) are pluripotent stem cells that can be established from dedifferentiation of all somatic cell types by epigenetic phenomena. iPSCs can be differentiated into any mature cells like neurons, hepatocytes, or pancreatic cells that have not been easily available to date. Thus, iPSCs are widely used for disease modeling, drug discovery, and cell therapy development. Here, we describe a protocol to obtain human mature and functional neutrophils and macrophages as ex vivo models of X-linked chronic granulomatous disease (X-CGD). This method can be applied to model the other genetic forms of CGD. We also describe methods for testing the characteristics and functions of neutrophils and macrophages by morphology, phagocytosis assay, release of granule markers or cytokines, cell surface markers, and NADPH oxidase activity.

Key words

iPSCs CGD Hematopoietic differentiation Neutrophils Macrophages NADPH oxidase Phagocytosis Exocytosis Cytokines 

Notes

Acknowledgments

MJS is grateful for the support from the University Grenoble Alpes (AGIR program 2014), the Faculty of Medicine and the Pole Recherche, University Hospital Grenoble Alpes, and Interreg France-Suisse (Programme de Cooperation Territoriale Europeenne, Fond Europeen de Developpement Regional (FEDER), 2017–2019). This work was also supported by the Delegation for Clinical Research and Innovations (DRCI, Rementips project 2014). We also thank Sylvain Beaumel and Michèle Mollin for their helpful and valuable work at the Centre Diagnostic et Recherche sur la CGD (CDiReC), Grenoble France. This article is dedicated to the memory of Cécile Martel, an outstanding technician at the CDiReC, who passed away recently. We miss you.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Julie Brault
    • 1
  • Bénédicte Vigne
    • 1
  • Marie José Stasia
    • 1
    • 2
    Email author
  1. 1.Centre Diagnostic et Recherche CGD (CDiReC), Pôle Biologie, CHU Grenoble AlpesGrenobleFrance
  2. 2.Universite Grenoble Alpes, CNRS, CEA, Institut de Biologie StructuraleGrenobleFrance

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