| Isolation, Culture, and Maintenance of Mouse Intestinal Stem Cells
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Author:
Date:
2016-02-20
[Abstract] In this protocol we describe our modifications to a method to isolate, culture and maintain mouse intestinal stem cells as crypt-villus forming organoids. These cells, isolated either from the small or large intestine, maintain self-renewal and multilineage differentiation potential over time. This provides investigators a tool to culture wild type or transformed intestinal epithelium, and a robust assay for stem cell tissue homeostasis in vitro.
[摘要] 在这个协议，我们描述我们修改分离，培养和维持鼠肠干细胞作为隐窝 - 绒毛形成类器官的方法。 从小肠或大肠分离的这些细胞随时间保持自我更新和多向分化潜能。 这为研究人员提供了培养野生型或转化的肠上皮的工具，以及用于体外干细胞组织稳态的稳健试验。
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| Isolation and 3-dimensional Culture of Primary Murine Intestinal Epithelial Cells
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Author:
Date:
2014-05-20
[Abstract] The intestine, together with skin and blood, belongs to the organs with the highest cell turnover, which makes it a perfect model to study cellular processes, such as proliferation and differentiation. Epithelial cell turnover in intestine is possible due to the presence of intestinal stem cells, which are located at the bottom of the crypt. Here, we recapitulate a detailed protocol for the isolation and culture procedures of primary epithelial intestinal cells in a three - dimensional (3D) in vitro system, described for the first time by Hans Clevers group (Sato et al., ...
[摘要] 肠道与皮肤和血液一起属于具有最高细胞更新的器官，这使得它成为研究细胞过程如增殖和分化的完美模型。由于存在位于隐窝底部的肠干细胞，肠上皮细胞更新是可能的。在这里，我们重述了在三维（3D）体外系统中原发性上皮肠细胞的分离和培养程序的详细方案，首次由Hans Clevers组（Sato > et al。，2009）。这种特定的3D培养物保留肠干细胞，其产生分化的子代，例如杯状细胞。该培养物具有许多应用并且代表研究干细胞生物学，上皮细胞再生和移植研究的有用模型。此外，呈现的3D文化可用于调查肠上皮细胞的屏障功能，以及上皮细胞和基质细胞之间的异型细胞相互作用。
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