| Neurosphere Co-culture Assay
|
|
Author:
Date:
2016-08-05
[Abstract] The hippocampal niche is one of two areas in the brain where stem cells reside. In this neurogenic niche, stem cells can be found in close proximity to astrocytes and in contact with microvessels consisting of pericytes and endothelial cells. To study the regulatory interplay of this complex niche network in a simplified in vitro model, we established a co-culture system. We investigate the formation of neurosphere under different co-culture conditions by using primary niche cells. Here, we describe the isolation procedure for primary niche cells culture of astrocytes, endothelial cells and pericytes/smooth muscle cells from mouse brain. These niche cells are co-cultured (by hanging inserts) with freshly isolated stem and precursor cells from the adult hippocampus to study the ...
[摘要] 真菌形态发生需要细胞壁的修饰和可塑性，这意味着其组分（包括壳多糖和葡聚糖）的合成和重塑。因此几丁质酶和葡聚糖酶活性对于细胞壁生物发生和细胞分裂是至关重要的。几丁质酶活性的定量可能有助于鉴定可能负面影响一些丝状真菌如产生细胞内和分泌型几丁质酶的尖孢镰刀菌的生长和形态发生的结构缺陷。基于它们对壳多糖底物的酶作用将壳聚糖分解酶分类。内酯酶被定义为催化几丁质链中内部点的随机裂解的酶。外切球蛋白酶催化乙酰壳寡糖或N-乙酰葡糖胺从几丁质的非还原端逐步释放，因此分别称为壳聚糖酶和β-N-乙酰氨基葡糖苷酶。在这里，我们描述了一种简单的方法来轻易地纯化几丁质酶，以便比较不同的F的内切几丁质酶活性和外切几丁质酶活性。尖孢镰孢菌株。该方案可适用于任何真菌物种。...
|
|
|
| Isolation of ILC2 from Mouse Liver
|
|
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。
|
|
|