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Specification of transplantable astroglial subtypes from human pluripotent stem cells

Nature Biotechnology volume 29, pages 528–534 (2011)Cite this article

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Abstract

Human pluripotent stem cells (hPSCs) have been differentiated efficiently to neuronal cell types. However, directed differentiation of hPSCs to astrocytes and astroglial subtypes remains elusive. In this study, hPSCs were directed to nearly uniform populations of immature astrocytes (>90% S100β+ and GFAP+) in large quantities. The immature human astrocytes exhibit similar gene expression patterns as primary astrocytes, display functional properties such as glutamate uptake and promotion of synaptogenesis, and become mature astrocytes by forming connections with blood vessels after transplantation into the mouse brain. Furthermore, hPSC-derived neuroepithelia, patterned to rostral-caudal and dorsal-ventral identities with the same morphogens used for neuronal subtype specification, generate immature astrocytes that express distinct homeodomain transcription factors and display phenotypic differences of different astroglial subtypes. These human astroglial progenitors and immature astrocytes will be useful for studying astrocytes in brain development and function, understanding the roles of astrocytes in disease processes and developing novel treatments for neurological disorders.

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Figure 1: Differentiation of astroglia from hPSCs.
Figure 2: Astroglial subtypes express region-specific proteins.
Figure 3: Functional characteristics of hPSC-derived immature astrocytes.
Figure 4: HPSC-derived astroglia retain their identity in vivo.
Figure 5: Hypothesis of astroglial subtype specification.

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Acknowledgements

The authors thank A. Messing for critical reading of the manuscript. This study was supported by the ALS Association, National Institute of Neurological Disorders and Stroke (NS045926, NS057778, NS064578), National MS Society (NMSS TR-3761), NYSTEM (C024406), Bleser Family Foundation, Busta Family Foundation, Neuroscience Training Program (T32 GM007507) and partly by a core grant to the Waisman Center from the National Institute of Child Health and Human Development (P30 HD03352).

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Author notes

  1. Jason P Weick and Yan Liu: These authors contributed equally to this work.

Authors and Affiliations

  1. Neuroscience Training Program, University of Wisconsin-Madison, Madison, Wisconsin, USA

    Robert Krencik & Su-Chun Zhang

  2. Waisman Center, University of Wisconsin–Madison, Madison, Wisconsin, USA

    Robert Krencik, Jason P Weick, Zhi-Jian Zhang & Su-Chun Zhang

  3. Department of Human Anatomy and Histology, Fudan University Shanghai Medical School, Shanghai, China

    Yan Liu & Su-Chun Zhang

  4. Department of Neuroscience, University of Wisconsin–Madison, Madison, Wisconsin, USA

    Su-Chun Zhang

  5. Department of Neurology, School of Medicine and Public Health, University of Wisconsin–Madison, Madison, Wisconsin, USA

    Su-Chun Zhang

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Contributions

R.K. and S.-C.Z. designed the experiments and wrote the manuscript. R.K., J.P.W., Y.L. and Z.-J.Z. performed the experiments. R.K., J.P.W., Y.L., Z.-.J.Z. and S.-C.Z. analyzed the data.

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Correspondence to Su-Chun Zhang.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Tables 1 and 2 and Supplementary Figures 1–4 (PDF 802 kb)

Supplementary Movie 1

Representative example of calcium wave propagation in FGF8-specified astroglia. (AVI 2201 kb)

Supplementary Movie 2

Z series of human astrocyte image in Fig. 4g stained for GFAP (green), human nuclei (red), and total nuclei (blue). (AVI 612 kb)

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Krencik, R., Weick, J., Liu, Y. et al. Specification of transplantable astroglial subtypes from human pluripotent stem cells. Nat Biotechnol 29, 528–534 (2011). https://doi.org/10.1038/nbt.1877

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