| Isolation and Culture of the Islets of Langerhans from Mouse Pancreas
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Author:
Date:
2016-06-20
[Abstract] The islets of Langerhans are clusters of endocrine cells located within the pancreas. Insulin-producing beta cells are the major cell type within islets, with glucagon-producing alpha cells and somatostatin-producing delta cells the other major cell types. The beta cells are the target of immune-mediated destruction in type 1 diabetes (Graham et al., 2012). Failure of beta cell function accompanied by loss of beta cell mass is also a feature of type 2 diabetes (Wali et al., 2013). Therefore studying the biology of pancreatic islets is important to understand the pathogenesis of diabetes and to develop new therapies. Here we describe the isolation of mouse islets. This requires gentle enzymatic and mechanical digestion of the exocrine tissue and density gradient ...
[摘要] 朗格汉斯岛是位于胰腺内的内分泌细胞簇。胰岛素生成β细胞是胰岛内的主要细胞类型，产生胰高血糖素的α细胞和生长抑素生成的δ细胞是其他主要细胞类型。 β细胞是免疫介导的1型糖尿病破坏的靶标（Graham等，2012）。 β细胞功能衰竭伴有β细胞量的丧失也是2型糖尿病的特征（Wali等，2013）。因此，研究胰岛的生物学对于了解糖尿病的发病机制和开发新的疗法是重要的。这里我们描述了小鼠胰岛的分离。这需要温和的酶促和机械消化外分泌组织和密度梯度分离（Chong et al。，2004; Liu和Shapiro，1995; Thomas et al。，1998）。然后，我们将描述胰岛如何可以整体培养或分散到单个细胞中，用于各种体外和体内分析。使用该协议可靠地导致200-400个胰岛的隔离，这取决于鼠标的应变。
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| Neutrophil Isolation from the Intestines
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Author:
Date:
2015-03-20
[Abstract] This protocol provides the possibility to isolate leukocytes including neutrophils out of intestinal tissues to use the received cells in further experiments of interest.
[摘要] 该方案提供了将白细胞包括嗜中性粒细胞从肠组织中分离的可能性，以在进一步的感兴趣的实验中使用接收的细胞。
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| Isolation of ILC2 from Mouse Liver
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Author:
Date:
2014-07-20
[Abstract] Group 2 innate lymphoid cells (ILC2) are a recently characterized cell population which lacks specific antigen receptors and contributes to immune responses at mucosal surfaces of lung and gut. Recently, we demonstrated that ILC2 expand in the context of chronic liver diseases in mice and contribute to pathology in an IL-33 dependent manner. Here, we describe a protocol to isolate highly purified ILC2 from mouse livers. This procedure provides effective digestion of liver tissue and limits proteolytic degradation of cell surface receptors leading to increased yields of biologically functional cells. The number of liver resident ILC2 in the steady state is low, however their number dramatically increase upon systemic or local treatment of mice with cytokines such as IL-25 and IL-33. Using ...
[摘要] 组2天然淋巴样细胞（ILC2）是最近表征的细胞群体，其缺乏特异性抗原受体并且有助于在肺和肠的粘膜表面的免疫应答。 最近，我们表明ILC2扩展在小鼠慢性肝病的背景下，并有助于病理学在IL-33依赖的方式。 在这里，我们描述了从小鼠肝脏分离高度纯化的ILC2的协议。 该方法提供肝组织的有效消化并限制细胞表面受体的蛋白水解降解，导致生物功能细胞的产量增加。 在稳定状态下肝脏驻留的ILC2的数量低，然而，在用细胞因子例如IL-25和IL-33全身或局部治疗小鼠时，它们的数量显着增加。 使用基于小环的表达构建体用于这些细胞因子，大量的功能性ILC2可以用本文提供的方案分离。
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