Generating Embryonic Salivary Gland Organoids.
Generating Embryonic Salivary Gland Organoids.
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DOI：
10.1002/cpcb.76
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发表时间：
2019-06
影响因子：
--
通讯作者：
Larsen M
中科院分区：
文献类型：
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作者：
Hosseini ZF;Nelson DA;Moskwa N;Larsen M
有机化合物是研究器官形态发生和分化的重要研究工具，因为它们在体外概括了细胞的自然三维组织，这对正常的细胞和器官功能是必不可少的。类有机物的组成可以被操纵以结合特定的细胞类型，以便于在类有机物形成期间对细胞-细胞相互作用的分子询问。我们描述了一种从胚胎唾液腺上皮祖细胞和间充质支持细胞中产生有机类化合物的方法。我们描述了从小鼠胚胎颌下腺中分离丰富的上皮细胞群作为簇，将间充质细胞作为单个细胞分离的方法。分离上皮细胞和间充质细胞群体允许对每种细胞类型进行独立的分子操作。此外，我们还描述了慢病毒转导间充质细胞的方法和有机物质的定量图像分析。这里描述的方法对于探索驱动器官形成的机制是有用的。
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. We describe a method for generating organoids derived from both embryonic salivary gland epithelial progenitor cells and mesenchymal support cells. We describe methods for isolating enriched populations of the epithelial cells as clusters and the mesenchyme cells as single cells from mouse embryonic submandibular salivary glands. Separating the epithelial and mesenchymal cell populations allows for independent molecular manipulation of each cell type. In addition, we describe methods for lentiviral transduction of the mesenchyme cells and quantitative image analysis of organoids. The methods described here are useful for exploring mechanisms driving organ formation.