Efficient conversion of human induced pluripotent stem cells into microglia by defined transcription factors

Shih-Wei Chen; Yu-Sheng Hung; Jong-Ling Fuh; Nien-Jung Chen; Yeh-Shiu Chu; Shu-Cian Chen; Ming-Ji Fann; Yu-Hui Wong

doi:10.1016/j.stemcr.2021.03.010

Efficient conversion of human induced pluripotent stem cells into microglia by defined transcription factors

Stem Cell Reports. 2021 May 11;16(5):1363-1380. doi: 10.1016/j.stemcr.2021.03.010. Epub 2021 Apr 8.

Authors

Shih-Wei Chen¹, Yu-Sheng Hung², Jong-Ling Fuh³, Nien-Jung Chen⁴, Yeh-Shiu Chu⁵, Shu-Cian Chen⁵, Ming-Ji Fann¹, Yu-Hui Wong⁶

Affiliations

¹ Brain Research Center, National Yang Ming Chiao Tung University, Taipei 112, Taiwan (ROC); Department of Life Sciences and Institute of Genome Sciences, School of Life Sciences, National Yang Ming Chiao Tung University, Taipei 112, Taiwan (ROC).
² Department of Life Sciences and Institute of Genome Sciences, School of Life Sciences, National Yang Ming Chiao Tung University, Taipei 112, Taiwan (ROC).
³ Brain Research Center, National Yang Ming Chiao Tung University, Taipei 112, Taiwan (ROC); Division of General Neurology, Neurological Institute, Taipei Veterans General Hospital, Taipei 112, Taiwan (ROC).
⁴ Institute of Microbiology and Immunology, School of Life Sciences, National Yang Ming Chiao Tung University, Taipei 112, Taiwan (ROC).
⁵ Brain Research Center, National Yang Ming Chiao Tung University, Taipei 112, Taiwan (ROC).
⁶ Brain Research Center, National Yang Ming Chiao Tung University, Taipei 112, Taiwan (ROC); Department of Life Sciences and Institute of Genome Sciences, School of Life Sciences, National Yang Ming Chiao Tung University, Taipei 112, Taiwan (ROC). Electronic address: yuhui.wong@nycu.edu.tw.

Abstract

Microglia, the immune cells of the central nervous system, play critical roles in brain physiology and pathology. We report a novel approach that produces, within 10 days, the differentiation of human induced pluripotent stem cells (hiPSCs) into microglia (iMG) by forced expression of both SPI1 and CEBPA. High-level expression of the main microglial markers and the purity of the iMG cells were confirmed by RT-qPCR, immunostaining, and flow cytometry analyses. Whole-transcriptome analysis demonstrated that these iMGs resemble human fetal/adult microglia but not human monocytes. Moreover, these iMGs exhibited appropriate physiological functions, including various inflammatory responses, ADP/ATP-evoked migration, and phagocytic ability. When co-cultured with hiPSC-derived neurons, the iMGs respond and migrate toward injured neurons. This study has established a protocol for the rapid conversion of hiPSCs into functional iMGs, which should facilitate functional studies of human microglia using different disease models and also help with drug discovery.

Keywords: CEBPA; SPI1/PU.1; induced microglia; induced pluripotent stem cells; reprogramming.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenosine Diphosphate / pharmacology
Adenosine Triphosphate / pharmacology
Biomarkers / metabolism
Calcium Signaling / drug effects
Cell Differentiation* / drug effects
Cell Differentiation* / genetics
Cell Movement / drug effects
Culture Media / pharmacology
Gene Expression Profiling
Gene Expression Regulation / drug effects
Humans
Induced Pluripotent Stem Cells / cytology*
Induced Pluripotent Stem Cells / drug effects
Induced Pluripotent Stem Cells / metabolism
Microglia / cytology*
Microglia / drug effects
Transcription Factors / metabolism*

Substances

Biomarkers
Culture Media
Transcription Factors
Adenosine Diphosphate
Adenosine Triphosphate