Directed Differentiation of Human Pluripotent Stem Cells to Microglia

Panagiotis Douvaras; Bruce Sun; Minghui Wang; Ilya Kruglikov; Gregory Lallos; Matthew Zimmer; Cecile Terrenoire; Bin Zhang; Sam Gandy; Eric Schadt; Donald O. Freytes; Scott Noggle; Valentina Fossati

doi:10.1016/j.stemcr.2017.04.023

Directed Differentiation of Human Pluripotent Stem Cells to Microglia

Panagiotis Douvaras
, Bruce Sun
, Minghui Wang
, Ilya Kruglikov
, Gregory Lallos
, Matthew Zimmer
, Cecile Terrenoire
, Bin Zhang
, Sam Gandy
, Eric Schadt
, Donald O. Freytes
, Scott Noggle
, Valentina Fossati

Research output: Contribution to journal › Article › peer-review

294 Scopus citations

Abstract

Microglia, the immune cells of the brain, are crucial to proper development and maintenance of the CNS, and their involvement in numerous neurological disorders is increasingly being recognized. To improve our understanding of human microglial biology, we devised a chemically defined protocol to generate human microglia from pluripotent stem cells. Myeloid progenitors expressing CD14/CX3CR1 were generated within 30 days of differentiation from both embryonic and induced pluripotent stem cells (iPSCs). Further differentiation of the progenitors resulted in ramified microglia with highly motile processes, expressing typical microglial markers. Analyses of gene expression and cytokine release showed close similarities between iPSC-derived (iPSC-MG) and human primary microglia as well as clear distinctions from macrophages. iPSC-MG were able to phagocytose and responded to ADP by producing intracellular Ca²⁺ transients, whereas macrophages lacked such response. The differentiation protocol was highly reproducible across several pluripotent stem cell lines.

Original language	English
Pages (from-to)	1516-1524
Number of pages	9
Journal	Stem Cell Reports
Volume	8
Issue number	6
DOIs	https://doi.org/10.1016/j.stemcr.2017.04.023
State	Published - 6 Jun 2017

Keywords

human microglia
human pluripotent stem cells
microglial differentiation

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10.1016/j.stemcr.2017.04.023

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title = "Directed Differentiation of Human Pluripotent Stem Cells to Microglia",

abstract = "Microglia, the immune cells of the brain, are crucial to proper development and maintenance of the CNS, and their involvement in numerous neurological disorders is increasingly being recognized. To improve our understanding of human microglial biology, we devised a chemically defined protocol to generate human microglia from pluripotent stem cells. Myeloid progenitors expressing CD14/CX3CR1 were generated within 30 days of differentiation from both embryonic and induced pluripotent stem cells (iPSCs). Further differentiation of the progenitors resulted in ramified microglia with highly motile processes, expressing typical microglial markers. Analyses of gene expression and cytokine release showed close similarities between iPSC-derived (iPSC-MG) and human primary microglia as well as clear distinctions from macrophages. iPSC-MG were able to phagocytose and responded to ADP by producing intracellular Ca2+ transients, whereas macrophages lacked such response. The differentiation protocol was highly reproducible across several pluripotent stem cell lines.",

keywords = "human microglia, human pluripotent stem cells, microglial differentiation",

author = "Panagiotis Douvaras and Bruce Sun and Minghui Wang and Ilya Kruglikov and Gregory Lallos and Matthew Zimmer and Cecile Terrenoire and Bin Zhang and Sam Gandy and Eric Schadt and Freytes, \{Donald O.\} and Scott Noggle and Valentina Fossati",

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year = "2017",

month = jun,

day = "6",

doi = "10.1016/j.stemcr.2017.04.023",

language = "English",

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TY - JOUR

T1 - Directed Differentiation of Human Pluripotent Stem Cells to Microglia

AU - Douvaras, Panagiotis

AU - Sun, Bruce

AU - Wang, Minghui

AU - Kruglikov, Ilya

AU - Lallos, Gregory

AU - Zimmer, Matthew

AU - Terrenoire, Cecile

AU - Zhang, Bin

AU - Gandy, Sam

AU - Schadt, Eric

AU - Freytes, Donald O.

AU - Noggle, Scott

AU - Fossati, Valentina

PY - 2017/6/6

Y1 - 2017/6/6

N2 - Microglia, the immune cells of the brain, are crucial to proper development and maintenance of the CNS, and their involvement in numerous neurological disorders is increasingly being recognized. To improve our understanding of human microglial biology, we devised a chemically defined protocol to generate human microglia from pluripotent stem cells. Myeloid progenitors expressing CD14/CX3CR1 were generated within 30 days of differentiation from both embryonic and induced pluripotent stem cells (iPSCs). Further differentiation of the progenitors resulted in ramified microglia with highly motile processes, expressing typical microglial markers. Analyses of gene expression and cytokine release showed close similarities between iPSC-derived (iPSC-MG) and human primary microglia as well as clear distinctions from macrophages. iPSC-MG were able to phagocytose and responded to ADP by producing intracellular Ca2+ transients, whereas macrophages lacked such response. The differentiation protocol was highly reproducible across several pluripotent stem cell lines.

AB - Microglia, the immune cells of the brain, are crucial to proper development and maintenance of the CNS, and their involvement in numerous neurological disorders is increasingly being recognized. To improve our understanding of human microglial biology, we devised a chemically defined protocol to generate human microglia from pluripotent stem cells. Myeloid progenitors expressing CD14/CX3CR1 were generated within 30 days of differentiation from both embryonic and induced pluripotent stem cells (iPSCs). Further differentiation of the progenitors resulted in ramified microglia with highly motile processes, expressing typical microglial markers. Analyses of gene expression and cytokine release showed close similarities between iPSC-derived (iPSC-MG) and human primary microglia as well as clear distinctions from macrophages. iPSC-MG were able to phagocytose and responded to ADP by producing intracellular Ca2+ transients, whereas macrophages lacked such response. The differentiation protocol was highly reproducible across several pluripotent stem cell lines.

KW - human microglia

KW - human pluripotent stem cells

KW - microglial differentiation

UR - https://www.scopus.com/pages/publications/85019390091

U2 - 10.1016/j.stemcr.2017.04.023

DO - 10.1016/j.stemcr.2017.04.023

M3 - Article

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AN - SCOPUS:85019390091

SN - 2213-6711

VL - 8

SP - 1516

EP - 1524

JO - Stem Cell Reports

JF - Stem Cell Reports

IS - 6

ER -

Directed Differentiation of Human Pluripotent Stem Cells to Microglia

Abstract

Keywords

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