A defined roadmap of radial glia and astrocyte differentiation from human pluripotent stem cells

Vukasin M. Jovanovic; Claire Weber; Jaroslav Slamecka; Seungmi Ryu; Pei Hsuan Chu; Chaitali Sen; Jason Inman; Juliana Ferreira De Sousa; Elena Barnaeva; Marissa Hirst; David Galbraith; Pinar Ormanoglu; Yogita Jethmalani; Jennifer Colon Mercado; Sam Michael; Michael E. Ward; Anton Simeonov; Ty C. Voss; Carlos A. Tristan; Ilyas Singeç

doi:10.1016/j.stemcr.2023.06.007

A defined roadmap of radial glia and astrocyte differentiation from human pluripotent stem cells

Vukasin M. Jovanovic, Claire Weber, Jaroslav Slamecka, Seungmi Ryu, Pei Hsuan Chu, Chaitali Sen, Jason Inman, Juliana Ferreira De Sousa, Elena Barnaeva, Marissa Hirst, David Galbraith, Pinar Ormanoglu, Yogita Jethmalani, Jennifer Colon Mercado, Sam Michael, Michael E. Ward, Anton Simeonov, Ty C. Voss, Carlos A. Tristan, Ilyas Singeç

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

15 Scopus citations

Abstract

Human gliogenesis remains poorly understood, and derivation of astrocytes from human pluripotent stem cells (hPSCs) is inefficient and cumbersome. Here, we report controlled glial differentiation from hPSCs that bypasses neurogenesis, which otherwise precedes astrogliogenesis during brain development and in vitro differentiation. hPSCs were first differentiated into radial glial cells (RGCs) resembling resident RGCs of the fetal telencephalon, and modulation of specific cell signaling pathways resulted in direct and stepwise induction of key astroglial markers (NFIA, NFIB, SOX9, CD44, S100B, glial fibrillary acidic protein [GFAP]). Transcriptomic and genome-wide epigenetic mapping and single-cell analysis confirmed RGC-to-astrocyte differentiation, obviating neurogenesis and the gliogenic switch. Detailed molecular and cellular characterization experiments uncovered new mechanisms and markers for human RGCs and astrocytes. In summary, establishment of a glia-exclusive neural lineage progression model serves as a unique serum-free platform of manufacturing large numbers of RGCs and astrocytes for neuroscience, disease modeling (e.g., Alexander disease), and regenerative medicine.

Original language	English
Pages (from-to)	1701-1720
Number of pages	20
Journal	Stem Cell Reports
Volume	18
Issue number	8
DOIs	https://doi.org/10.1016/j.stemcr.2023.06.007
State	Published - 8 Aug 2023
Externally published	Yes

Keywords

Alexander disease
CEPT cocktail
astrocytes
dual SMAD inhibition
gliogenesis
gliogenic switch
iPSCs
multi-omics
neural differentiation
radial glia
robotic cell culture
serum-free
single-cell analysis

ASJC Scopus subject areas

Biochemistry
Genetics
Developmental Biology
Cell Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.stemcr.2023.06.007

Cite this

Jovanovic, V. M., Weber, C., Slamecka, J., Ryu, S., Chu, P. H., Sen, C., Inman, J., De Sousa, J. F., Barnaeva, E., Hirst, M., Galbraith, D., Ormanoglu, P., Jethmalani, Y., Mercado, J. C., Michael, S., Ward, M. E., Simeonov, A., Voss, T. C., Tristan, C. A., & Singeç, I. (2023). A defined roadmap of radial glia and astrocyte differentiation from human pluripotent stem cells. Stem Cell Reports, 18(8), 1701-1720. https://doi.org/10.1016/j.stemcr.2023.06.007

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title = "A defined roadmap of radial glia and astrocyte differentiation from human pluripotent stem cells",

abstract = "Human gliogenesis remains poorly understood, and derivation of astrocytes from human pluripotent stem cells (hPSCs) is inefficient and cumbersome. Here, we report controlled glial differentiation from hPSCs that bypasses neurogenesis, which otherwise precedes astrogliogenesis during brain development and in vitro differentiation. hPSCs were first differentiated into radial glial cells (RGCs) resembling resident RGCs of the fetal telencephalon, and modulation of specific cell signaling pathways resulted in direct and stepwise induction of key astroglial markers (NFIA, NFIB, SOX9, CD44, S100B, glial fibrillary acidic protein [GFAP]). Transcriptomic and genome-wide epigenetic mapping and single-cell analysis confirmed RGC-to-astrocyte differentiation, obviating neurogenesis and the gliogenic switch. Detailed molecular and cellular characterization experiments uncovered new mechanisms and markers for human RGCs and astrocytes. In summary, establishment of a glia-exclusive neural lineage progression model serves as a unique serum-free platform of manufacturing large numbers of RGCs and astrocytes for neuroscience, disease modeling (e.g., Alexander disease), and regenerative medicine.",

keywords = "Alexander disease, CEPT cocktail, astrocytes, dual SMAD inhibition, gliogenesis, gliogenic switch, iPSCs, multi-omics, neural differentiation, radial glia, robotic cell culture, serum-free, single-cell analysis",

author = "Jovanovic, {Vukasin M.} and Claire Weber and Jaroslav Slamecka and Seungmi Ryu and Chu, {Pei Hsuan} and Chaitali Sen and Jason Inman and {De Sousa}, {Juliana Ferreira} and Elena Barnaeva and Marissa Hirst and David Galbraith and Pinar Ormanoglu and Yogita Jethmalani and Mercado, {Jennifer Colon} and Sam Michael and Ward, {Michael E.} and Anton Simeonov and Voss, {Ty C.} and Tristan, {Carlos A.} and Ilyas Singe{\c c}",

note = "Publisher Copyright: {\textcopyright} 2023",

year = "2023",

month = aug,

day = "8",

doi = "10.1016/j.stemcr.2023.06.007",

language = "English",

volume = "18",

pages = "1701--1720",

journal = "Stem Cell Reports",

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Jovanovic, VM, Weber, C, Slamecka, J, Ryu, S, Chu, PH, Sen, C, Inman, J, De Sousa, JF, Barnaeva, E, Hirst, M, Galbraith, D, Ormanoglu, P, Jethmalani, Y, Mercado, JC, Michael, S, Ward, ME, Simeonov, A, Voss, TC, Tristan, CA & Singeç, I 2023, 'A defined roadmap of radial glia and astrocyte differentiation from human pluripotent stem cells', Stem Cell Reports, vol. 18, no. 8, pp. 1701-1720. https://doi.org/10.1016/j.stemcr.2023.06.007

TY - JOUR

T1 - A defined roadmap of radial glia and astrocyte differentiation from human pluripotent stem cells

AU - Jovanovic, Vukasin M.

AU - Weber, Claire

AU - Slamecka, Jaroslav

AU - Ryu, Seungmi

AU - Chu, Pei Hsuan

AU - Sen, Chaitali

AU - Inman, Jason

AU - De Sousa, Juliana Ferreira

AU - Barnaeva, Elena

AU - Hirst, Marissa

AU - Galbraith, David

AU - Ormanoglu, Pinar

AU - Jethmalani, Yogita

AU - Mercado, Jennifer Colon

AU - Michael, Sam

AU - Ward, Michael E.

AU - Simeonov, Anton

AU - Voss, Ty C.

AU - Tristan, Carlos A.

AU - Singeç, Ilyas

PY - 2023/8/8

Y1 - 2023/8/8

N2 - Human gliogenesis remains poorly understood, and derivation of astrocytes from human pluripotent stem cells (hPSCs) is inefficient and cumbersome. Here, we report controlled glial differentiation from hPSCs that bypasses neurogenesis, which otherwise precedes astrogliogenesis during brain development and in vitro differentiation. hPSCs were first differentiated into radial glial cells (RGCs) resembling resident RGCs of the fetal telencephalon, and modulation of specific cell signaling pathways resulted in direct and stepwise induction of key astroglial markers (NFIA, NFIB, SOX9, CD44, S100B, glial fibrillary acidic protein [GFAP]). Transcriptomic and genome-wide epigenetic mapping and single-cell analysis confirmed RGC-to-astrocyte differentiation, obviating neurogenesis and the gliogenic switch. Detailed molecular and cellular characterization experiments uncovered new mechanisms and markers for human RGCs and astrocytes. In summary, establishment of a glia-exclusive neural lineage progression model serves as a unique serum-free platform of manufacturing large numbers of RGCs and astrocytes for neuroscience, disease modeling (e.g., Alexander disease), and regenerative medicine.

AB - Human gliogenesis remains poorly understood, and derivation of astrocytes from human pluripotent stem cells (hPSCs) is inefficient and cumbersome. Here, we report controlled glial differentiation from hPSCs that bypasses neurogenesis, which otherwise precedes astrogliogenesis during brain development and in vitro differentiation. hPSCs were first differentiated into radial glial cells (RGCs) resembling resident RGCs of the fetal telencephalon, and modulation of specific cell signaling pathways resulted in direct and stepwise induction of key astroglial markers (NFIA, NFIB, SOX9, CD44, S100B, glial fibrillary acidic protein [GFAP]). Transcriptomic and genome-wide epigenetic mapping and single-cell analysis confirmed RGC-to-astrocyte differentiation, obviating neurogenesis and the gliogenic switch. Detailed molecular and cellular characterization experiments uncovered new mechanisms and markers for human RGCs and astrocytes. In summary, establishment of a glia-exclusive neural lineage progression model serves as a unique serum-free platform of manufacturing large numbers of RGCs and astrocytes for neuroscience, disease modeling (e.g., Alexander disease), and regenerative medicine.

KW - Alexander disease

KW - CEPT cocktail

KW - astrocytes

KW - dual SMAD inhibition

KW - gliogenesis

KW - gliogenic switch

KW - iPSCs

KW - multi-omics

KW - neural differentiation

KW - radial glia

KW - robotic cell culture

KW - serum-free

KW - single-cell analysis

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U2 - 10.1016/j.stemcr.2023.06.007

DO - 10.1016/j.stemcr.2023.06.007

M3 - Article

C2 - 37451260

AN - SCOPUS:85166666718

SN - 2213-6711

VL - 18

SP - 1701

EP - 1720

JO - Stem Cell Reports

JF - Stem Cell Reports

IS - 8

ER -

A defined roadmap of radial glia and astrocyte differentiation from human pluripotent stem cells

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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