Single-cell transcriptomics defines an improved, validated monoculture protocol for differentiation of human iPSC to microglia
Abstract There is increasing genetic evidence for the role of microglia in neurodegenerative diseases, including Alzheimer’s, Parkinson’s, and motor neuron disease. Therefore, there is a need to generate authentic in vitro models to study human microglial physiology. Various methods have been develo...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2022-11-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-23477-2 |
| _version_ | 1798016902811877376 |
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| author | Sam J. Washer Marta Perez-Alcantara Yixi Chen Juliette Steer William S. James Gosia Trynka Andrew R. Bassett Sally A. Cowley |
| author_facet | Sam J. Washer Marta Perez-Alcantara Yixi Chen Juliette Steer William S. James Gosia Trynka Andrew R. Bassett Sally A. Cowley |
| author_sort | Sam J. Washer |
| collection | DOAJ |
| description | Abstract There is increasing genetic evidence for the role of microglia in neurodegenerative diseases, including Alzheimer’s, Parkinson’s, and motor neuron disease. Therefore, there is a need to generate authentic in vitro models to study human microglial physiology. Various methods have been developed using human induced Pluripotent Stem Cells (iPSC) to generate microglia, however, systematic approaches to identify which media components are actually essential for functional microglia are mostly lacking. Here, we systematically assess medium components, coatings, and growth factors required for iPSC differentiation to microglia. Using single-cell RNA sequencing, qPCR, and functional assays, with validation across two labs, we have identified several medium components from previous protocols that are redundant and do not contribute to microglial identity. We provide an optimised, defined medium which produces both transcriptionally and functionally relevant microglia for modelling microglial physiology in neuroinflammation and for drug discovery. |
| first_indexed | 2024-04-11T15:56:55Z |
| format | Article |
| id | doaj.art-8437721023204dd8b1fa075d4b5d56b5 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-11T15:56:55Z |
| publishDate | 2022-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-8437721023204dd8b1fa075d4b5d56b52022-12-22T04:15:07ZengNature PortfolioScientific Reports2045-23222022-11-0112111610.1038/s41598-022-23477-2Single-cell transcriptomics defines an improved, validated monoculture protocol for differentiation of human iPSC to microgliaSam J. Washer0Marta Perez-Alcantara1Yixi Chen2Juliette Steer3William S. James4Gosia Trynka5Andrew R. Bassett6Sally A. Cowley7James and Lillian Martin Centre for Stem Cell Research, Sir William Dunn School of Pathology, University of OxfordWellcome Sanger Institute, Wellcome Genome CampusWellcome Sanger Institute, Wellcome Genome CampusWellcome Sanger Institute, Wellcome Genome CampusJames and Lillian Martin Centre for Stem Cell Research, Sir William Dunn School of Pathology, University of OxfordWellcome Sanger Institute, Wellcome Genome CampusWellcome Sanger Institute, Wellcome Genome CampusJames and Lillian Martin Centre for Stem Cell Research, Sir William Dunn School of Pathology, University of OxfordAbstract There is increasing genetic evidence for the role of microglia in neurodegenerative diseases, including Alzheimer’s, Parkinson’s, and motor neuron disease. Therefore, there is a need to generate authentic in vitro models to study human microglial physiology. Various methods have been developed using human induced Pluripotent Stem Cells (iPSC) to generate microglia, however, systematic approaches to identify which media components are actually essential for functional microglia are mostly lacking. Here, we systematically assess medium components, coatings, and growth factors required for iPSC differentiation to microglia. Using single-cell RNA sequencing, qPCR, and functional assays, with validation across two labs, we have identified several medium components from previous protocols that are redundant and do not contribute to microglial identity. We provide an optimised, defined medium which produces both transcriptionally and functionally relevant microglia for modelling microglial physiology in neuroinflammation and for drug discovery.https://doi.org/10.1038/s41598-022-23477-2 |
| spellingShingle | Sam J. Washer Marta Perez-Alcantara Yixi Chen Juliette Steer William S. James Gosia Trynka Andrew R. Bassett Sally A. Cowley Single-cell transcriptomics defines an improved, validated monoculture protocol for differentiation of human iPSC to microglia Scientific Reports |
| title | Single-cell transcriptomics defines an improved, validated monoculture protocol for differentiation of human iPSC to microglia |
| title_full | Single-cell transcriptomics defines an improved, validated monoculture protocol for differentiation of human iPSC to microglia |
| title_fullStr | Single-cell transcriptomics defines an improved, validated monoculture protocol for differentiation of human iPSC to microglia |
| title_full_unstemmed | Single-cell transcriptomics defines an improved, validated monoculture protocol for differentiation of human iPSC to microglia |
| title_short | Single-cell transcriptomics defines an improved, validated monoculture protocol for differentiation of human iPSC to microglia |
| title_sort | single cell transcriptomics defines an improved validated monoculture protocol for differentiation of human ipsc to microglia |
| url | https://doi.org/10.1038/s41598-022-23477-2 |
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