Cargando…
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...
Autores principales: | , , , , , , , |
---|---|
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 |
_version_ | 1784708049798168576 |
---|---|
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. |
format | Online Article Text |
id | pubmed-9105476 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT lanferjonas efficientandeasyconversionofhumanipscsintofunctionalinducedmicroglialikecells AT kaindljohanna efficientandeasyconversionofhumanipscsintofunctionalinducedmicroglialikecells AT krummlaura efficientandeasyconversionofhumanipscsintofunctionalinducedmicroglialikecells AT gonzalezaceramiguel efficientandeasyconversionofhumanipscsintofunctionalinducedmicroglialikecells AT neurathmarkus efficientandeasyconversionofhumanipscsintofunctionalinducedmicroglialikecells AT regensburgermartin efficientandeasyconversionofhumanipscsintofunctionalinducedmicroglialikecells AT krachflorian efficientandeasyconversionofhumanipscsintofunctionalinducedmicroglialikecells AT winnerbeate efficientandeasyconversionofhumanipscsintofunctionalinducedmicroglialikecells |