Robust induction of functional astrocytes using NGN2 expression in human pluripotent stem cells

Martin H Berryer; Matthew Tegtmeyer; Loïc Binan; Vera Valakh; Anna Nathanson; Darina Trendafilova; Ethan Crouse; Jenny A Klein; Daniel Meyer; Olli Pietiläinen; Francesca Rapino; Samouil L Farhi; Lee L Rubin; Steven A McCarroll; Ralda Nehme; Lindy E Barrett

doi:10.1016/j.isci.2023.106995

Robust induction of functional astrocytes using NGN2 expression in human pluripotent stem cells

iScience. 2023 May 30;26(7):106995. doi: 10.1016/j.isci.2023.106995. eCollection 2023 Jul 21.

Authors

Martin H Berryer^{1

2}, Matthew Tegtmeyer^{1

2

3}, Loïc Binan^{1

4}, Vera Valakh^{1

4}, Anna Nathanson^{1

2}, Darina Trendafilova^{1

2}, Ethan Crouse^{1

2}, Jenny A Klein^{1

2}, Daniel Meyer¹, Olli Pietiläinen^{1

5}, Francesca Rapino², Samouil L Farhi^{1

4}, Lee L Rubin², Steven A McCarroll^{1

6}, Ralda Nehme^{1

2}, Lindy E Barrett^{1

2}

Affiliations

¹ Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
² Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA.
³ Centre for Gene Therapy and Regenerative Medicine, King's College, London, UK.
⁴ Klarman Cell Observatory, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
⁵ University of Helsinki, Helsinki, Finland.
⁶ Department of Genetics, Harvard Medical School, Boston, MA, USA.

Abstract

Emerging evidence of species divergent features of astrocytes coupled with the relative inaccessibility of human brain tissue underscore the utility of human pluripotent stem cell (hPSC) technologies for the generation and study of human astrocytes. However, existing approaches for hPSC-astrocyte generation are typically lengthy or require intermediate purification steps. Here, we establish a rapid and highly scalable method for generating functional human induced astrocytes (hiAs). These hiAs express canonical astrocyte markers, respond to pro-inflammatory stimuli, exhibit ATP-induced calcium transients and support neuronal network development. Moreover, single-cell transcriptomic analyses reveal the generation of highly reproducible cell populations across individual donors, mostly resembling human fetal astrocytes. Finally, hiAs generated from a trisomy 21 disease model identify expected alterations in cell-cell adhesion and synaptic signaling, supporting their utility for disease modeling applications. Thus, hiAs provide a valuable and practical resource for the study of basic human astrocyte function and dysfunction in disease.

Keywords: Methodology in biological sciences; Neuroscience; Transcriptomics.

Abstract

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