期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 23, 期 9, 页码 -出版社
MDPI
DOI: 10.3390/ijms23094526
关键词
microglia; pluripotent stem cells; cell differentiation; neurology; immunology
资金
- Bavarian Ministry of Science and the Arts in the framework of the ForInter network
- Interdisciplinary Center for Clinical Research (IZKF) at the University Hospital Erlangen [P054, J88, A5 (2019-02)]
- German Research Foundation [DFG WI 3567/2-1, 270949263/GRK2162]
- IZKF advanced project [E30]
- TreatHSP consortium (BMBF) [01GM1905B]
In this study, we developed an easy-to-use method to convert human induced pluripotent stem cells into functional microglia-like cells. The protocol involved efficient differentiation and maturation steps, sequential harvesting and freezing techniques, as well as culturing on uncoated glass surfaces to increase yield while maintaining differentiation efficiency and functional characteristics. This method is highly valuable for research on 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 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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