| Whole-mount Enteroid Proliferation Staining
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Author:
Date:
2016-06-20
[Abstract] Small intestinal organoids, otherwise known as enteroids, have become an increasingly utilized model for intestinal biology in vitro as they recapitulate the various epithelial cells within the intestinal crypt (Mahe et al., 2013; Sato et al., 2009). Assessment of growth dynamics within these cultures is an important step to understanding how alterations in gene expression, treatment with protective and toxic agents, and genetic mutations alter properties essential for crypt growth and survival as well as the stem cell properties of the individual cells within the crypt. This protocol describes a method of visualization of proliferating cells within the crypt in three dimensions (Barrett et al., 2015). Whole-mount proliferation staining of enteroids ...
[摘要] 小肠类器官，也称为肠袢，已经成为越来越多地用于肠道生物学的体外模型，因为它们重现了在肠隐窝内的各种上皮细胞（Mahe等人 ，2013; Sato et al。，2009）。在这些培养物中评估生长动力学是理解基因表达的改变，用保护性和毒性试剂处理以及遗传突变改变隐窝生长和存活所必需的性质以及细胞内单个细胞的干细胞性质的重要步骤地穴。该方案描述了在隐窝内三维的增殖细胞的可视化方法（Barrett等人，2015）。使用EdU掺入的肠衣的整体增殖染色使得研究者能够观察到肠内的所有增殖细胞，而不是如在包埋和切片中所见到的在薄切片中获得生长信息，确保来自干细胞区室的增殖的真实表现到隐窝的终末分化细胞。
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| Mouse ESC Differentiation to Nkx2.1+ Lung and Thyroid Progenitors
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Author:
Date:
2012-11-20
[Abstract] The de novo derivation of lung progenitors from pluripotent stem cells provides the opportunity to model early lung development in vitro and allows easy access to cells for tissue engineering or basic cell biology studies. This detailed protocol allows the generation of lung and thyroid progenitors from mouse embryonic stem cell (ESC) or induced pluripotent stem cell (iPSC) lines. When used together with a published Nkx2.1-GFP knock-in ESC line, the protocol allows tracking and purification of lung and thyroid progenitors by sorting on the GFP reporter based on the induction of the earliest known marker of lung and thyroid cell fate, Nkx2.1. After sorting, a pure population of Nkx2.1+ cells can then be replated for further expansion, differentiation, and maturation in ...
[摘要] 来自多能干细胞的肺祖细胞的新发现提供了在体外模拟早期肺发育的机会，并且容易获得用于组织工程或基础细胞生物学研究的细胞。 该详细方案允许从小鼠胚胎干细胞（ESC）或诱导多能干细胞（iPSC）系生成肺和甲状腺祖细胞。 当与公布的Nkx2.1-GFP敲入ESC系一起使用时，该方案允许通过基于对最早已知的肺和甲状腺细胞命运的标记物的诱导在GFP报告基因上进行分选来跟踪和纯化肺和甲状腺祖细胞， Nkx2.1。 在分选后，然后可以在无血清条件下培养纯化的Nkx2.1 +细胞群，以进一步扩增，分化和成熟。
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