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NFIA is a gliogenic switch enabling rapid derivation of functional human astrocytes from pluripotent stem cells


The mechanistic basis of gliogenesis, which occurs late in human development, is poorly understood. Here we identify nuclear factor IA (NFIA) as a molecular switch inducing human glial competency. Transient expression of NFIA is sufficient to trigger glial competency of human pluripotent stem cell-derived neural stem cells within 5 days and to convert these cells into astrocytes in the presence of glial-promoting factors, as compared to 3–6 months using current protocols. NFIA-induced astrocytes promote synaptogenesis, exhibit neuroprotective properties, display calcium transients in response to appropriate stimuli and engraft in the adult mouse brain. Differentiation involves rapid but reversible chromatin remodeling, glial fibrillary acidic protein (GFAP) promoter demethylation and a striking lengthening of the G1 cell cycle phase. Genetic or pharmacological manipulation of G1 length partially mimics NFIA function. We used the approach to generate astrocytes with region-specific or reactive features. Our study defines key mechanisms of the gliogenic switch and enables the rapid production of human astrocytes for disease modeling and regenerative medicine.

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Fig. 1: Transient expression of NFIA in neuroepithelial stem cells confers glial competency.
Fig. 2: NFIA-induced astrocytes are functional.
Fig. 3: NFIA cannot maintain the glial-competent state.
Fig. 4: NFIA expression leads to a slower G1 cell cycle phase to induce glial competency.

Data availability

The data and reagents in this study are available from the corresponding author upon reasonable request. All FASTQ files and Supplementary files were uploaded to National Center for Biotechnology Information Gene Expression Omnibus under accession code GSE104232.


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We are grateful to the members the Studer laboratory for helpful discussions and support for this project, and to G. Cederquist, M. Tomishima, S. Irion and V. Tabar for their critical comments on the manuscript. We would also like to give special thanks to A. Koff (MSKCC) for his helpful comments, experimental discussions regarding the cell cycle and comments on the manuscript. Additionally, we would like to thank A. Viale at Integrated Genomics Operation Core (MSKCC) for the RNA-sequencing studies, M. Witkin at the Epigenetics Core (MSKCC) for the ATAC sequencing, S. Fujisawa, E. Feng and V. Boyko at the Molecular Cytology Core (MSKCC) for help in calcium imaging studies and quantification of synaptic proteins, R. Garripa and H. Liu at the RNAi Core (MSKCC) for help with short hairpin RNA design and the Flow Cytometry Core (MSKCC) for the cell-sorting applications. J.T. was supported by the Tri-I Starr Stem Cell Scholars postdoctoral training fellowship. S.R.G was supported by the Ruth L. Kirschstein Individual Predoctoral NRSA for MD/PhD Fellowship (No. 1F30MH115616-01) and by a Medical Scientist Training Program grant from the National Institute of General Medical Sciences of the National Institutes of Health (No. T32GM007739) to the Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD Program. E.M.G. was supported by a grant from the Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (No. 323630-164217). The work was supported by grants to L.S. from the National Institutes of Health (No. R21NS084334, No. R01AG056298), by core grant No. P30CA008748 and by a grant from Project ALS.

Author information




J.T. contributed to the conception, study design, data analysis and interpretation, writing of the manuscript, bioinformatics, development and execution of directed differentiation strategies from hPSCs, generation of LTNSCs, cell cycle analysis and calcium imaging. E.L.C. contributed to maintenance of hPSCs and directed differentiation of spinal cord progenitors. S.R.G. contributed to cell cycle analysis and astrocyte activation assays. E.M.G.contributed to transplantation studies and data analysis. K.A.A. and P.A.G. contributed to electrophysiology and assessment of neuronal maturation. J.A.S. contributed to generation of LTNSCs and calcium imaging. L.S. contributed to the conception, study design, data analysis and interpretation and writing of the manuscript.

Corresponding authors

Correspondence to Jason Tchieu or Lorenz Studer.

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Competing interests

The Memorial Sloan-Kettering Cancer Center has filed a patent application (WO2018175574A1) on the methods described in the manuscript. L.S. is the scientific cofounder of Bluerock Therapeutics.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–25

Reporting Summary

Supplementary Table 1

Gene expression dataset of NFIA-induced astrocytes log transformed counts per million (CPM)

Supplementary Table 2

Gene expression dataset of the NFIA-induction time course. Fold changes compared to day 0 (neurogenic NSCs)

Supplementary Table 3

List of antibodies used in this study

Supplementary Video 1

FUCCI-O reporter on LTNSCs without doxycycline induction

Supplementary Video 2

FUCCI-O reporter on LTNSCs with continuous doxycycline treatment

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Tchieu, J., Calder, E.L., Guttikonda, S.R. et al. NFIA is a gliogenic switch enabling rapid derivation of functional human astrocytes from pluripotent stem cells. Nat Biotechnol 37, 267–275 (2019).

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