Chapter title |
Generating Inner Ear Organoids from Mouse Embryonic Stem Cells.
|
---|---|
Chapter number | 215 |
Book title |
Embryonic Stem Cell Protocols
|
Published in |
Methods in molecular biology, March 2015
|
DOI | 10.1007/7651_2015_215 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2953-5, 978-1-4939-2954-2
|
Authors |
Longworth-Mills, Emma, Koehler, Karl R, Hashino, Eri, Emma Longworth-Mills, Karl R. Koehler, Eri Hashino, Koehler, Karl R. |
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. |
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