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Directed Differentiation of Mouse Embryonic Stem Cells Into Inner Ear Sensory Epithelia in 3D Culture

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Organ Regeneration

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1597))

Abstract

The inner ear sensory epithelium harbors mechanosensory hair cells responsible for detecting sound and maintaining balance. This protocol describes a three-dimensional (3D) culture system that efficiently generates inner ear sensory epithelia from aggregates of mouse embryonic stem (mES) cells. By mimicking the activations and repressions of key signaling pathways during in vivo inner ear development, mES cell aggregates are sequentially treated with recombinant proteins and small molecule inhibitors for activating or inhibiting the Bmp, TGFβ, Fgf, and Wnt signaling pathways. These stepwise treatments promote mES cells to sequentially differentiate into epithelia representing the non-neural ectoderm, preplacodal ectoderm, otic placodal ectoderm, and ultimately, the hair cell-containing sensory epithelia. The derived hair cells are surrounded by a layer of supporting cells and are innervated by sensory neurons. This in vitro inner ear organoid culture system may serve as a valuable tool in developmental and physiological research, disease modeling, drug testing, and potential cell-based therapies.

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Acknowledgments

This work was supported by a National Institutes of Health grant R01 DC013294 (to E.H.).

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Authors and Affiliations

  1. Department of Otolaryngology—Head and Neck Surgery, and Stark Neurosciences Research Institute, Indiana University School of Medicine, 980 W. Walnut St., Indianapolis, IN, 46202, USA

    Jing Nie, Karl R. Koehler & Eri Hashino

Authors
  1. Jing Nie
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  2. Karl R. Koehler
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  3. Eri Hashino
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Corresponding author

Correspondence to Eri Hashino .

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Editors and Affiliations

  1. Laboratory for Organ Regeneration, RIKEN Center for Developmental Biology, Kobe, Hyogo, Japan

    Takashi Tsuji

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Nie, J., Koehler, K.R., Hashino, E. (2017). Directed Differentiation of Mouse Embryonic Stem Cells Into Inner Ear Sensory Epithelia in 3D Culture. In: Tsuji, T. (eds) Organ Regeneration. Methods in Molecular Biology, vol 1597. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6949-4_6

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  • DOI: https://doi.org/10.1007/978-1-4939-6949-4_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6947-0

  • Online ISBN: 978-1-4939-6949-4

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