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Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells

Current protocols converting human induced pluripotent stem cells (iPSCs) into induced microglia-like cells (iMGL) are either dependent on overexpression of transcription factors or require substantial experience in stem-cell technologies. Here, we developed an easy-to-use two-step protocol to conve...

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Autores principales: Lanfer, Jonas, Kaindl, Johanna, Krumm, Laura, Gonzalez Acera, Miguel, Neurath, Markus, Regensburger, Martin, Krach, Florian, Winner, Beate
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9105476/
https://www.ncbi.nlm.nih.gov/pubmed/35562917
http://dx.doi.org/10.3390/ijms23094526
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author Lanfer, Jonas
Kaindl, Johanna
Krumm, Laura
Gonzalez Acera, Miguel
Neurath, Markus
Regensburger, Martin
Krach, Florian
Winner, Beate
author_facet Lanfer, Jonas
Kaindl, Johanna
Krumm, Laura
Gonzalez Acera, Miguel
Neurath, Markus
Regensburger, Martin
Krach, Florian
Winner, Beate
author_sort Lanfer, Jonas
collection PubMed
description Current protocols converting human induced pluripotent stem cells (iPSCs) into induced microglia-like cells (iMGL) are either dependent on overexpression of transcription factors or require substantial experience in stem-cell technologies. Here, we developed an easy-to-use two-step protocol to convert iPSCs into functional iMGL via: (1) highly efficient differentiation of hematopoietic progenitor cells (HPCs) from iPSCs, and (2) optimized maturation of HPCs to iMGL. A sequential harvesting approach led to an increased HPC yield. The protocol implemented a freezing step, thus allowing HPC biobanking and flexible timing of differentiation into iMGL. Our iMGL responded adequately to the inflammatory stimuli LPS, and iMGL RNAseq analysis matched those of other frequently used protocols. Comparing three different coating modalities, we increased the iMGL yield by culturing on uncoated glass surfaces, thereby retaining differentiation efficiency and functional hallmarks of iMGL. In summary, we provide a high-quality, easy-to-use protocol, rendering generation and functional studies on iMGL an accessible lab resource.
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spelling pubmed-91054762022-05-14 Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells Lanfer, Jonas Kaindl, Johanna Krumm, Laura Gonzalez Acera, Miguel Neurath, Markus Regensburger, Martin Krach, Florian Winner, Beate Int J Mol Sci Article Current protocols converting human induced pluripotent stem cells (iPSCs) into induced microglia-like cells (iMGL) are either dependent on overexpression of transcription factors or require substantial experience in stem-cell technologies. Here, we developed an easy-to-use two-step protocol to convert iPSCs into functional iMGL via: (1) highly efficient differentiation of hematopoietic progenitor cells (HPCs) from iPSCs, and (2) optimized maturation of HPCs to iMGL. A sequential harvesting approach led to an increased HPC yield. The protocol implemented a freezing step, thus allowing HPC biobanking and flexible timing of differentiation into iMGL. Our iMGL responded adequately to the inflammatory stimuli LPS, and iMGL RNAseq analysis matched those of other frequently used protocols. Comparing three different coating modalities, we increased the iMGL yield by culturing on uncoated glass surfaces, thereby retaining differentiation efficiency and functional hallmarks of iMGL. In summary, we provide a high-quality, easy-to-use protocol, rendering generation and functional studies on iMGL an accessible lab resource. MDPI 2022-04-20 /pmc/articles/PMC9105476/ /pubmed/35562917 http://dx.doi.org/10.3390/ijms23094526 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lanfer, Jonas
Kaindl, Johanna
Krumm, Laura
Gonzalez Acera, Miguel
Neurath, Markus
Regensburger, Martin
Krach, Florian
Winner, Beate
Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells
title Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells
title_full Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells
title_fullStr Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells
title_full_unstemmed Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells
title_short Efficient and Easy Conversion of Human iPSCs into Functional Induced Microglia-like Cells
title_sort efficient and easy conversion of human ipscs into functional induced microglia-like cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9105476/
https://www.ncbi.nlm.nih.gov/pubmed/35562917
http://dx.doi.org/10.3390/ijms23094526
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