Generating Embryonic Salivary Gland Organoids

Q3 Biochemistry, Genetics and Molecular Biology

Current Protocols in Cell Biology Pub Date : 2018-11-05 DOI:10.1002/cpcb.76

Zeinab F. Hosseini, Deirdre A. Nelson, Nicholas Moskwa, Melinda Larsen

{"title":"Generating Embryonic Salivary Gland Organoids","authors":"Zeinab F. Hosseini, Deirdre A. Nelson, Nicholas Moskwa, Melinda Larsen","doi":"10.1002/cpcb.76","DOIUrl":null,"url":null,"abstract":"Organoids are important research tools for studying organ morphogenesis and differentiation because they recapitulate ex vivo the native 3D organization of cells that is essential for proper cell and organ function. The composition of organoids can be manipulated to incorporate specific cell types to facilitate molecular interrogation of cell‐cell interactions during organoid formation. A method for generating organoids derived from both embryonic salivary gland epithelial progenitor cells and mesenchymal support cells is described. Methods for isolating enriched populations of the epithelial cells as clusters and the mesenchyme cells as single cells from mouse embryonic submandibular salivary glands are also provided. Separating the epithelial and mesenchymal cell populations allows for independent molecular manipulation of each cell type. In addition, methods for lentiviral transduction of the mesenchyme cells and quantitative image analysis of organoids are provided. The methods described here are useful for exploring mechanisms driving organ formation. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":40051,"journal":{"name":"Current Protocols in Cell Biology","volume":"83 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpcb.76","citationCount":"11","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Cell Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpcb.76","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}

引用次数: 11

Abstract

Organoids are important research tools for studying organ morphogenesis and differentiation because they recapitulate ex vivo the native 3D organization of cells that is essential for proper cell and organ function. The composition of organoids can be manipulated to incorporate specific cell types to facilitate molecular interrogation of cell‐cell interactions during organoid formation. A method for generating organoids derived from both embryonic salivary gland epithelial progenitor cells and mesenchymal support cells is described. Methods for isolating enriched populations of the epithelial cells as clusters and the mesenchyme cells as single cells from mouse embryonic submandibular salivary glands are also provided. Separating the epithelial and mesenchymal cell populations allows for independent molecular manipulation of each cell type. In addition, methods for lentiviral transduction of the mesenchyme cells and quantitative image analysis of organoids are provided. The methods described here are useful for exploring mechanisms driving organ formation. © 2018 by John Wiley & Sons, Inc.

Abstract Image

查看原文本刊更多论文

产生胚胎唾液腺类器官

类器官是研究器官形态发生和分化的重要研究工具，因为它们概括了体外细胞的天然三维组织，这是正常细胞和器官功能所必需的。类器官的组成可以被操纵，以纳入特定的细胞类型，以促进类器官形成过程中细胞间相互作用的分子询问。描述了一种从胚胎唾液腺上皮祖细胞和间充质支持细胞生成类器官的方法。本文还提供了从小鼠胚胎颌下唾液腺中分离成簇的上皮细胞和单细胞的间充质细胞的方法。分离上皮细胞和间充质细胞群允许对每种细胞类型进行独立的分子操作。此外，还提供了间充质细胞慢病毒转导和类器官定量图像分析的方法。这里描述的方法对于探索驱动器官形成的机制是有用的。©2018 by John Wiley &儿子,Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Current Protocols in Cell Biology Biochemistry, Genetics and Molecular Biology-Cell Biology

自引率

0.00%

发文量

期刊介绍： Developed by leading scientists in the field, Current Protocols in Cell Biology is an essential reference for researchers who study the relationship between specific molecules and genes and their location, function and structure at the cellular level. Updated every three months in all formats, CPCB is constantly evolving to keep pace with the very latest discoveries and developments.