小鼠耳蜗细胞分选及细胞类型特异性类器官培养。

Murine cochlear cell sorting and cell-type-specific organoid culture.

作者信息

Kubota Marie, Heller Stefan

机构信息

Department of Otolaryngology - Head & Neck Surgery, Stanford University School of Medicine, Stanford, CA 94305, USA.

Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.

出版信息

STAR Protoc. 2021 Jul 7;2(3):100645. doi: 10.1016/j.xpro.2021.100645. eCollection 2021 Sep 17.

DOI:10.1016/j.xpro.2021.100645

PMID:34278332

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8271165/

Abstract

Neonatal mouse cochlear duct cells can proliferate and grow into inner ear organoids. Distinctive cochlear duct cell types have different organoid formation capacities. Here, we provide a flow cytometric cell-sorting method that allows the subsequent culture of individual cochlear cell populations. For the efficient culture of the sorted cells, we provide protocols for growing free-floating inner ear organoids, the adherence of organoids to a substrate, and the expansion of organoid-derived inner ear colonies. For complete details on the use and execution of this protocol, please refer to Kubota et al. (2021).

摘要

新生小鼠耳蜗管细胞可以增殖并生长为内耳类器官。不同类型的耳蜗管细胞具有不同的类器官形成能力。在这里，我们提供了一种流式细胞术细胞分选方法，可用于后续对单个耳蜗细胞群体进行培养。为了有效培养分选后的细胞，我们提供了培养自由漂浮内耳类器官、使类器官附着于基质以及扩增类器官衍生的内耳集落的方案。有关本方案使用和实施的完整详细信息，请参考久保田等人（2021年）的研究。