| Mouse CD8+ T Cell Migration in vitro and CXCR3 Internalization Assays
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Author:
Date:
2017-03-20
[Abstract] Chemokines are molecules that regulate the positioning of cells during homeostasis and inflammation. CXCL10 is an interferon-induced chemokine that attracts cells that express the chemokine receptor CXCR3 on their surface. CXCL10 expression is often induced upon inflammation and guides lymphocytes, such as T and NK cells, into the injured tissues. Notably, CXCL10 binding to CXCR3 induces receptor internalization and, therefore, low CXCR3 levels in cells positive for CXCR3 expression can be indicative of chemokine signaling.
Here, we describe an in vitro method to evaluate the ability of murine CD8+ T cells to migrate towards recombinant murine CXCL10; and a flow cytometry assay to measure CXCR3 expression levels at the surface of T cells, after exposure to ...
[摘要] 趋化因子是调节体内平衡和炎症期间细胞定位的分子。 CXCL10是干扰素诱导的趋化因子，其吸引在其表面上表达趋化因子受体CXCR3的细胞。 CXCL10表达通常在炎症诱导并引导淋巴细胞如T和NK细胞进入受损组织。值得注意的是，CXCL10与CXCR3结合诱导受体内化，因此CXCR3表达阳性细胞中的CXCR3水平降低可能是趋化因子信号传导的指示。
＆nbsp;这里，我们描述体外方法来评估鼠CD8 + T细胞向重组鼠CXCL10迁移的能力;以及暴露于不同剂量的趋化因子后在T细胞表面测量CXCR3表达水平的流式细胞术测定。
背景 趋化因子介导的T细胞运输是稳态和炎症期间的重要过程。活化的CD8 + T细胞表达趋化因子受体，例如CXCR3，允许它们向趋化因子CXCL9,10和11迁移，通常在损伤组织上上调。调节T细胞迁移的分子线索的评估对于了解其功能背后的生物学非常重要，但是在体内运行的复杂机制有时难以去卷积。在这里，我们提供有关体外方法的详细信息，以评估CD8 + T细胞上的趋化因子功能，重点是CXCL10介导的化学吸引和CXCR3内化。我们使用可以容易地在体外扩增和活化的抗原特异性转基因CD8 +细胞，因此提供足够数量的表型相同的淋巴细胞（例如， ...
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| Organotypic Spinal Cord Slice Cultures and a Method to Detect Cell Proliferation in These Slices
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Author:
Date:
2016-10-05
[Abstract] In these culture models, the normal cytoarchitecture and local neuronal circuits of the spinal cord are preserved, offering a compromise between dissociated cell cultures and complete animal studies. The addition of 5-ethynyl-2’-deoxyuridine (EdU) to the culture medium allows for the detection of proliferating cells.
[摘要] 在这些文化模型中，保留了脊髓的正常细胞结构和局部神经元回路，提供了解离的细胞培养物和完全动物研究之间的折衷。 向培养基中加入5-乙炔基-2'-脱氧尿苷（EdU）允许检测增殖细胞。
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| Mouse Subependymal Zone Explants Cultured on Primary Astrocytes
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Author:
Date:
2016-07-20
[Abstract] Neuroblast migration is a major component in the process of neuronal circuit assembly. In the rodent postnatal brain, the subependymal zone (SEZ) is the largest neurogenic niche where new neurons are born throughout life. These cells migrate several millimeters following a specific pathway called the rostral migratory stream (RMS) towards the olfactory bulb. Once they reach their final destination, they differentiate mainly as interneurons and integrate into already established neuronal circuits. Along the RMS, neuroblasts use a particular mode of migration known as chain migration. They stay attached to each other while migrating and are intimately associated with adjacent astrocytic processes. To dissect the molecular and cellular signals involved in neuroblast migration, we developed ...
[摘要] 神经母细胞迁移是神经元电路组装过程中的主要组成部分。 在啮齿动物出生后的大脑中，子宫颈区（SEZ）是新生神经元生命中最大的神经源性生态位。 这些细胞沿着称为传播流（RMS）的特定途径朝向嗅球迁移几毫米。 一旦达到其最终目的地，它们主要区分为中间神经元并整合到已建立的神经元回路中。 沿着RMS，神经母细胞使用称为链转移的特定迁移模式。 它们在迁移时保持相互依赖，并与相邻的星形细胞过程密切相关。 为了解剖神经母细胞迁移中涉及的分子和细胞信号，我们开发了一种体外系统，其由在星形胶质细胞单层顶部共培养的经SEZ衍生的外植体组成。
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