Directed differentiation of mouse embryonic stem cells into inner ear sensory epithelia in 3D culture

Jing Nie, Karl R. Koehler, Eri Hashino

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages67-83
Number of pages17
DOIs
StatePublished - Jan 1 2017

Publication series

NameMethods in Molecular Biology
Volume1597
ISSN (Print)1064-3745

Keywords

  • Hair cells
  • Inner ear
  • Mouse pluripotent stem cells
  • Organoid
  • Sensory epithelium
  • Three-dimensional culture
  • Vestibular

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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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 Methods in Molecular Biology (pp. 67-83). (Methods in Molecular Biology; Vol. 1597). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6949-4_6