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Generation of pure monocultures of human microglia-like cells from induced pluripotent stem cells
- Source :
- Banerjee, P, Paza, E, Perkins, E M, James, O G, Kenkhuis, B, Lloyd, A F, Burr, K, Story, D, Yusuf, D, He, X, Backofen, R, Dando, O, Chandran, S & Priller, J 2020, ' Generation of pure monocultures of human microglia-like cells from induced pluripotent stem cells ', Stem Cell Research, vol. 49, 102046 . https://doi.org/10.1016/j.scr.2020.102046, Stem Cell Research, Vol 49, Iss, Pp 102046-(2020), Stem cell research 49, 102046 (2020). doi:10.1016/j.scr.2020.102046
- Publication Year :
- 2020
-
Abstract
- Microglia are resident tissue macrophages of the central nervous system (CNS) that arise from erythromyeloid progenitors during embryonic development. They play essential roles in CNS development, homeostasis and response to disease. Since microglia are difficult to procure from the human brain, several protocols have been developed to generate microglia-like cells from human induced pluripotent stem cells (hiPSCs). However, some concerns remain over the purity and quality of in vitro generated microglia. Here, we describe a new protocol that does not require co-culture with neural cells and yields cultures of 100% P2Y12+ 95% TMEM119+ ramified human microglia-like cells (hiPSC-MG). In the presence of neural precursor cell-conditioned media, hiPSC-MG expressed high levels of human microglia signature genes, including SALL1, CSF1R, P2RY12, TMEM119, TREM2, HEXB and SIGLEC11, as revealed by whole-transcriptome analysis. Stimulation of hiPSC-MG with lipopolysaccharide resulted in downregulation of P2Y12 expression, induction of IL1B mRNA expression and increase in cell capacitance. HiPSC-MG were phagocytically active and maintained their cell identity after transplantation into murine brain slices and human brain spheroids. Together, our new protocol for the generation of microglia-like cells from human iPSCs will facilitate the study of human microglial function in health and disease.
- Subjects :
- 0301 basic medicine
Nervous system
Organoid
organoid
Induced Pluripotent Stem Cells
microglia
Biology
03 medical and health sciences
Mice
0302 clinical medicine
Trem2 protein, mouse
Downregulation and upregulation
ddc:570
medicine
Animals
Humans
Progenitor cell
Receptors, Immunologic
Induced pluripotent stem cell
lcsh:QH301-705.5
Neurons
Membrane Glycoproteins
Microglia
TREM2 protein, human
TREM2
nervous system
Brain
Cell Biology
General Medicine
Human brain
Cell biology
Transplantation
030104 developmental biology
medicine.anatomical_structure
lcsh:Biology (General)
myeloid cells
Myeloid cells
Transcriptome
transcriptome
030217 neurology & neurosurgery
Developmental Biology
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Banerjee, P, Paza, E, Perkins, E M, James, O G, Kenkhuis, B, Lloyd, A F, Burr, K, Story, D, Yusuf, D, He, X, Backofen, R, Dando, O, Chandran, S & Priller, J 2020, ' Generation of pure monocultures of human microglia-like cells from induced pluripotent stem cells ', Stem Cell Research, vol. 49, 102046 . https://doi.org/10.1016/j.scr.2020.102046, Stem Cell Research, Vol 49, Iss, Pp 102046-(2020), Stem cell research 49, 102046 (2020). doi:10.1016/j.scr.2020.102046
- Accession number :
- edsair.doi.dedup.....45a7ad277ce5fb4e57e907ecbb752ab1