| Detection of Cell Death in Planarians
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Author:
Date:
2018-10-05
[Abstract] Planarians are freshwater flatworms, well known for their ability to regenerate a complete organism from any piece of their body. Furthermore, planarians are constantly growing and degrowing throughout their lives, maintaining a functional and proportioned body. These properties rely on the presence of a population of adult stem cells and on the tight control of their cell renewal, which is based on the balance between the proliferation of new cells and their differentiation, and the death of unnecessary cells. Due to the importance of these two processes in planarian biology, over the years, researchers have optimized molecular techniques to detect both cell proliferation and cell death in planarians. Here, we present the two main protocols currently used for cell death detection and ...
[摘要] 涡虫是淡水扁虫，因其能够从身体的任何部分再生完整的有机体而闻名。 此外，涡虫在其一生中不断生长和去除，保持功能和比例的身体。 这些特性依赖于成体干细胞群的存在以及对细胞更新的严格控制，其基于新细胞增殖与其分化之间的平衡以及不必要细胞的死亡。 由于这两个过程在涡虫生物学中的重要性，多年来，研究人员已经优化了分子技术，以检测涡虫中的细胞增殖和细胞死亡。 在这里，我们提出了目前用于细胞死亡检测和量化的两种主要方案：Caspase-3活性定量和TUNEL分析。
【背景】成体生物体中的细胞更新是基于三个过程的复杂机制：（a）通过细胞死亡消除选定的细胞; （b）通过细胞分裂取代已消除的细胞，通常涉及成体干细胞及其后代; （c）新生细胞的分化及其与先前存在的组织的整合（Pellettieri和Sanchez ...
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| Enhancement of Mucus Production in Eukaryotic Cells and Quantification of Adherent Mucus by ELISA
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Author:
Date:
2018-06-20
[Abstract] The mucosal surfaces of the gastrointestinal, respiratory, reproductive, and urinary tracts, and the surface of the eye harbor a resident microflora that lives in symbiosis with their host and forms a complex ecosystem. The protection of the vulnerable epithelium is primarily achieved by mucins that form a gel-like structure adherent to the apical cell surface. This mucus layer constitutes a physical and chemical barrier between the microbial flora and the underlying epithelium. Mucus is critical to the maintenance of a homeostatic relationship between the microbiota and its host. Subtle deviations from this dynamic interaction may result in major implications for health. The protocol in this article describes the procedures to grow low mucus-producing HT29 and high mucus-producing ...
[摘要] 胃肠道，呼吸道，生殖道和泌尿道的粘膜表面以及眼睛表面都有一个居住的微生物群落，它们与宿主共生并形成一个复杂的生态系统。脆弱的上皮细胞的保护主要通过形成附着于顶端细胞表面的凝胶样结构的粘蛋白实现。该粘液层构成了微生物菌群和下层上皮之间的物理和化学屏障。粘液对维持微生物群与宿主之间的稳态关系至关重要。与这种动态互动的细微差异可能会对健康产生重大影响。本文中的方案描述了生长低粘液产生HT29和高粘液产生HT29-MTX-E12细胞的程序，维持细胞并通过ELISA将其用于粘液定量。此外，还介绍了如何评估分泌黏液的数量。该系统可用于研究粘液对抗细菌毒素的保护作用，例如测试不同培养条件对粘液产生的影响或分析分子通过粘液层的扩散。由于本方案中使用的ELISA可用于不同的物种和粘液蛋白，因此也可以使用其他细胞类型。
【背景】身体与外部环境的界面由粘膜表面形成。这些粘膜上皮组织可以在例如胃肠道，呼吸道，生殖道和尿道以及眼睛表面发现。由于它们暴露于外部环境中，许多微生物会聚集在这些组织中。因此，这些上皮细胞已经进化出多种防御机制来回应其易受微生物攻击的影响。许多防御性化合物被分泌到粘膜液中，包括粘蛋白，抗体，防御素，protegrin，聚集蛋白，cathelicidins，溶菌酶，组蛋白和一氧化氮（Kagnoff和Eckmann，1997，Lu等人，2002 ...
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| 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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