Generation of Human Neutrophils from Induced Pluripotent Stem Cells in Chemically Defined Conditions Using ETV2 Modified mRNA

This protocol describes a rapid and efficient feeder-, serum-, and xeno-free method for neutrophil generation from hiPSCs using ETV2 modified mRNA (mmRNA), which directs hematoendothelial programming of hiPSCs. Hematoendothelial progenitors were cultured with GM-CSF, FGF-2, and UM171 to expand myelo...

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Autores principales: Majumder, Aditi, Suknuntha, Kran, Bennin, David, Klemm, Lucas, Brok-Volchanskaya, Vera S., Huttenlocher, Anna, Slukvin, Igor
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7543976/
https://www.ncbi.nlm.nih.gov/pubmed/33043305
http://dx.doi.org/10.1016/j.xpro.2020.100075
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author Majumder, Aditi
Suknuntha, Kran
Bennin, David
Klemm, Lucas
Brok-Volchanskaya, Vera S.
Huttenlocher, Anna
Slukvin, Igor
author_facet Majumder, Aditi
Suknuntha, Kran
Bennin, David
Klemm, Lucas
Brok-Volchanskaya, Vera S.
Huttenlocher, Anna
Slukvin, Igor
author_sort Majumder, Aditi
collection PubMed
description This protocol describes a rapid and efficient feeder-, serum-, and xeno-free method for neutrophil generation from hiPSCs using ETV2 modified mRNA (mmRNA), which directs hematoendothelial programming of hiPSCs. Hematoendothelial progenitors were cultured with GM-CSF, FGF-2, and UM171 to expand myelomonocytic progenitors, followed by treatment with G-CSF and retinoic acid agonist Am580 to induce neutrophil maturation. This protocol is suitable for generating functional neutrophils from iPSCs to interrogate the role of genes in a neutrophil development and function. For complete details on the use and execution of this protocol, please refer to Brok-Volchanskaya et al. (2019).
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spelling pubmed-75439762020-10-08 Generation of Human Neutrophils from Induced Pluripotent Stem Cells in Chemically Defined Conditions Using ETV2 Modified mRNA Majumder, Aditi Suknuntha, Kran Bennin, David Klemm, Lucas Brok-Volchanskaya, Vera S. Huttenlocher, Anna Slukvin, Igor STAR Protoc Protocol This protocol describes a rapid and efficient feeder-, serum-, and xeno-free method for neutrophil generation from hiPSCs using ETV2 modified mRNA (mmRNA), which directs hematoendothelial programming of hiPSCs. Hematoendothelial progenitors were cultured with GM-CSF, FGF-2, and UM171 to expand myelomonocytic progenitors, followed by treatment with G-CSF and retinoic acid agonist Am580 to induce neutrophil maturation. This protocol is suitable for generating functional neutrophils from iPSCs to interrogate the role of genes in a neutrophil development and function. For complete details on the use and execution of this protocol, please refer to Brok-Volchanskaya et al. (2019). Elsevier 2020-07-27 /pmc/articles/PMC7543976/ /pubmed/33043305 http://dx.doi.org/10.1016/j.xpro.2020.100075 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Majumder, Aditi
Suknuntha, Kran
Bennin, David
Klemm, Lucas
Brok-Volchanskaya, Vera S.
Huttenlocher, Anna
Slukvin, Igor
Generation of Human Neutrophils from Induced Pluripotent Stem Cells in Chemically Defined Conditions Using ETV2 Modified mRNA
title Generation of Human Neutrophils from Induced Pluripotent Stem Cells in Chemically Defined Conditions Using ETV2 Modified mRNA
title_full Generation of Human Neutrophils from Induced Pluripotent Stem Cells in Chemically Defined Conditions Using ETV2 Modified mRNA
title_fullStr Generation of Human Neutrophils from Induced Pluripotent Stem Cells in Chemically Defined Conditions Using ETV2 Modified mRNA
title_full_unstemmed Generation of Human Neutrophils from Induced Pluripotent Stem Cells in Chemically Defined Conditions Using ETV2 Modified mRNA
title_short Generation of Human Neutrophils from Induced Pluripotent Stem Cells in Chemically Defined Conditions Using ETV2 Modified mRNA
title_sort generation of human neutrophils from induced pluripotent stem cells in chemically defined conditions using etv2 modified mrna
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7543976/
https://www.ncbi.nlm.nih.gov/pubmed/33043305
http://dx.doi.org/10.1016/j.xpro.2020.100075
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