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.
CITATION STYLE
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). Humana Press Inc. https://doi.org/10.1007/7651_2015_215
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