Generating inner ear organoids from mouse embryonic stem cells

Emma Longworth-Mills, Karl R. Koehler, Eri Hashino

Research output: Chapter in Book/Report/Conference proceedingChapter

13 Citations (Scopus)

Abstract

This protocol describes a three-dimensional culture method for generating inner ear sensory epithelia, which comprises sensory hair cells and a concurrently arising neuronal population. Mouse embryonic stem cells are initially plated in 96-well plates with differentiation media; following aggregation, Matrigel is added in order to promote epithelialization. A series of small molecule applications is then used over the first 14 days of culture to guide differentiation towards an otic lineage. After 16-20 days, vesicles containing inner ear sensory hair cells and supporting cells arise from the cultured aggregates. Aggregates may be analyzed using immunohistochemistry and electrophysiology techniques. This system serves as a simple and relatively inexpensive in vitro model of inner ear development.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages391-406
Number of pages16
Volume1341
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1341
ISSN (Print)10643745

Fingerprint

Organoids
Inner Ear
Electrophysiology
Ear
Cultured Cells
Epithelium
Immunohistochemistry
Population
Mouse Embryonic Stem Cells

Keywords

  • Cell cultureTechniques
  • Cell differentiation
  • Hair cell
  • Inner ear
  • Neurogenesis
  • Stem cell
  • Three-dimensional cell culture
  • Vestibular

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Longworth-Mills, E., Koehler, K. R., & Hashino, E. (2016). Generating inner ear organoids from mouse embryonic stem cells. In Methods in Molecular Biology (Vol. 1341, pp. 391-406). (Methods in Molecular Biology; Vol. 1341). Humana Press Inc.. https://doi.org/10.1007/7651_2015_215

Generating inner ear organoids from mouse embryonic stem cells. / Longworth-Mills, Emma; Koehler, Karl R.; Hashino, Eri.

Methods in Molecular Biology. Vol. 1341 Humana Press Inc., 2016. p. 391-406 (Methods in Molecular Biology; Vol. 1341).

Research output: Chapter in Book/Report/Conference proceedingChapter

Longworth-Mills, E, Koehler, KR & Hashino, E 2016, Generating inner ear organoids from mouse embryonic stem cells. in Methods in Molecular Biology. vol. 1341, Methods in Molecular Biology, vol. 1341, Humana Press Inc., pp. 391-406. https://doi.org/10.1007/7651_2015_215
Longworth-Mills E, Koehler KR, Hashino E. Generating inner ear organoids from mouse embryonic stem cells. In Methods in Molecular Biology. Vol. 1341. Humana Press Inc. 2016. p. 391-406. (Methods in Molecular Biology). https://doi.org/10.1007/7651_2015_215
Longworth-Mills, Emma ; Koehler, Karl R. ; Hashino, Eri. / Generating inner ear organoids from mouse embryonic stem cells. Methods in Molecular Biology. Vol. 1341 Humana Press Inc., 2016. pp. 391-406 (Methods in Molecular Biology).
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