3D mouse embryonic stem cell culture for generating inner ear organoids.
3D mouse embryonic stem cell culture for generating inner ear organoids.
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DOI：
10.1038/nprot.2014.100
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发表时间：
2014
期刊：
影响因子：
14.8
通讯作者：
Hashino E
中科院分区：
文献类型：
--
作者：
Koehler KR;Hashino E
This protocol describes a culture system in which inner ear sensory tissue is produced from mouse embryonic stem cells under chemically defined conditions. This model is amenable to basic and translational investigations into inner ear biology and regeneration. In this protocol, mouse embryonic stem cells are aggregated in 96-well plates in medium containing extracellular matrix proteins to promote epithelialization. During the first 14 days a series of precisely timed protein and small molecule treatments sequentially induce epithelia that represent the mouse embryonic nonneural ectoderm, preplacodal ectoderm and otic vesicle epithelia. Ultimately, these tissues develop into cysts with a pseudostratified epithelium containing inner ear hair cells and supporting cells after 16–20 days. Concurrently, sensory-like neurons generate synapse-like structures with the derived hair cells. We have designated the stem cell-derived epithelia harboring hair cells, supporting cells and sensory-like neurons inner ear organoids. This method provides a reproducible and scalable means to generate inner ear sensory tissue in vitro.
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