| Adoptive Transfer of Lung Antigen Presenting Cells
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Author:
Date:
2017-03-20
[Abstract] Our protocol describes adoptive transfer of antigen presenting cells (APCs) isolated from the lungs by enzymatic digestion and magnetic enrichment. This protocol can be used to study APC functions and trafficking.
[摘要] 我们的方案描述了通过酶消化和磁力富集从肺分离的抗原呈递细胞（APC）的过继转移。该协议可用于研究APC功能和贩运。
背景 包括巨噬细胞和树突状细胞（DC）在内的肺癌APC在感染侵袭性病原体，引发T细胞应答和控制耐受性反应中发挥关键作用。肺DC是稳定状态下最有效的专业APC，包括常规DC（cDC）和浆细胞样DC（pDC），以及炎症时新招募的单核细胞衍生的DC（moDC）（Kopf等人）。 ，2015）。肺静脉巨噬细胞，包括肺泡巨噬细胞，间质巨噬细胞和支气管巨噬细胞，在呈递抗原方面的效力较低。
＆nbsp;所有巨噬细胞和cDC在其细胞表面表达高水平的CD11c，而pDC表达中等水平的CD11c（Becher等，2014）。因此，CD11c磁性微珠可用于分离小鼠肺巨噬细胞和cDC。我们开发了一种从正常小鼠中分离肺APCs的方案，然后将其过渡转移到同基因骨髓移植小鼠。我们使用该方案来确定正常的肺APCs是否足以恢复移植后疱疹病毒感染的T辅助细胞极化（Zhou等人，2016）。
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| Neurosphere Co-culture Assay
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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。...
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| Isolating Taste Buds and Taste Cells from Vallate Papillae of C57BL/6J Mice for Detecting Transmitter Secretion
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Author:
Date:
2016-06-05
[Abstract] Mouse is a well-accepted model for studying taste bud function. Mice readily detect and respond to taste substances that humans consider to have sweet, bitter, salty, sour and umami taste qualities. A great deal of recent research on taste receptors is based on this species. Live mice are needed for these experiments because no alternative in vitro model incorporates all elements of taste transduction and peripheral signaling. The C57BL/6J strain was selected because these mice respond robustly to many taste stimuli and because of variety of transgenic animals, such as PLCβ2-GFP and GAD67-GFP, were derived from that strain. Prior analyses on behavior, nerve responses, cellular electrophysiology and molecular biology, all conducted on C57BL/6J mice will form a solid foundation for ...
[摘要] 鼠标是研究味蕾功能的一个被广泛接受的模式。小鼠容易检测和响应人们认为具有甜味，苦味，咸味，酸味和鲜味的品质的味道。最近对味觉感受器的大量研究是基于这个物种。这些实验需要活的小鼠，因为没有替代的体外模型包含所有的味觉转导和外周信号的元件。选择C57BL / 6J菌株，因为这些小鼠对许多味觉刺激反应强烈，并且由于来自该菌株的多种转基因动物，例如PLCβ2-GFP和GAD67-GFP。对C57BL / 6J小鼠进行的行为，神经反应，细胞电生理学和分子生物学的先前分析将为拟议研究奠定坚实的基础（Finger et al。，2005; Huang and Wu，2015; Huang et al。，2007 ）。因此，新鲜安乐死的动物必须用作味蕾的来源，我们将从中分离出味蕾和味道细胞。
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