Directed Differentiation of Human Pluripotent Stem Cells into Inner Ear Organoids.
Directed Differentiation of Human Pluripotent Stem Cells into Inner Ear Organoids.
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人类多能干细胞定向分化为内耳类器官。
DOI：
10.1007/7651_2021_448
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Hashino,Eri
中科院分区：
文献类型：
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作者：
Ueda,Yoshitomo;Moore,StephenT;Hashino,Eri
内耳的感觉上皮细胞含有机械敏感的毛细胞，可以检测声音和头部加速度。该方案详细描述了一种3D分化方法，以产生含有感觉上皮细胞和毛细胞的内耳类器官。将人多能干细胞聚集在低结合96孔板中，并在具有细胞外基质的化学成分确定的培养基中处理以促进上皮形成。小分子和重组蛋白以逐步的方式应用，以概括体内内耳发育期间存在的形态发生线索（BMP、TGF-β、FGF和WNT）。这些治疗诱导的顺序形成的非神经外胚层，耳上鳃祖域，和耳基板。衍生的耳基板然后经历自我引导的形态发生以形成耳囊，其最终产生含有毛细胞和支持细胞的感觉上皮，以及具有与毛细胞的突触形成的神经元。这种人干细胞来源的内耳类器官系统为在体外研究人内耳发育和疾病提供了理想的平台。
The sensory epithelia of the inner ear contain mechanosensitive hair cells that detect sound and head acceleration. This protocol details a 3D differentiation method to generate inner ear organoids containing sensory epithelia with hair cells. Human pluripotent stem cells are aggregated in low-binding 96-well plates and treated in chemically defined media with extracellular matrix to promote epithelialization. Small molecules and recombinant proteins are applied in a stepwise manner to recapitulate the morphogenic cues (BMP, TGF-β, FGF, and WNT) present during inner ear development in vivo. These treatments induce the sequential formation of nonneural ectoderm, otic-epibranchial progenitor domain, and otic placodes. The derived otic placodes then undergo self-guided morphogenesis to form otic vesicles, which eventually give rise to sensory epithelia containing hair cells and supporting cells, as well as neurons with synaptic formations to hair cells. This human stem cell–derived inner ear organoid system provides an ideal platform to study human inner ear development and disease in vitro.