| Sacral Spinal Cord Transection and Isolated Sacral Cord Preparation to Study Chronic Spinal Cord Injury in Adult Mice
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Author:
Date:
2018-04-05
[Abstract] Spinal cord injury (SCI) is characterized by multiple sensory/motor impairments that arise from different underlying neural mechanisms. Linking specific sensory/motor impairments to neural mechanism is limited by a lack of direct experimental access to these neural circuits. Here, we describe an experimental model which addresses this shortcoming. We generated a mouse model of chronic spinal cord injury that reliably reproduces spasticity observed after SCI, while at the same time allows study of motor impairments in vivo and in an in vitro preparation of the spinal cord. The model allows for the combination of mouse genetics in in vitro and in vivo conditions with advanced imaging, behavioral analysis, and detailed electrophysiology, techniques which ...
[摘要] 脊髓损伤（SCI）的特点是由不同的潜在神经机制引起的多种感觉/运动损伤。 将特定的感觉/运动障碍与神经机制联系起来受限于缺乏对这些神经回路的直接实验访问。 在这里，我们描述了一个解决这个缺点的实验模型。 我们产生了一种慢性脊髓损伤的小鼠模型，其可靠地再现SCI后观察到的痉挛状态，同时允许研究运动损伤体内和体外制剂 的脊髓。 该模型允许将体外和体内条件下的小鼠遗传与高级成像，行为分析和详细的电生理学技术相结合，这些技术在常规SCI中不易应用 楷模。
【背景】脊髓损伤导致了毁灭性的感官 - ...
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| Immune Cell Isolation from Mouse Femur Bone Marrow
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Author:
Date:
2015-10-20
[Abstract] The bone marrow is the site of hematopoiesis and contains mixed population of blood cells including erythrocytes, granulocytes, monocytes, dendritic cells, lymphocytes and hematopoietic stem cells. The following protocol provides a simple and fast method for isolation of bone marrow immune cells (no erythrocytes) from mouse femurs with a yield of approximate 8 x 107 cells in 5 ml culture media (1.6 x 104 cells/µl). Further isolation or flow cytometric analysis might be required for study of specific immune cell types.
[摘要] 骨髓是造血的部位，包含红细胞，粒细胞，单核细胞，树突状细胞，淋巴细胞和造血干细胞等混合血细胞。 以下方案提供了一种简单而快速的方法，用于在5ml培养基（1.6×10 4个细胞/μl）中分离来自小鼠股骨的骨髓免疫细胞（无红细胞），产量约为8×10 7个细胞。 研究特异性免疫细胞类型可能需要进一步的分离或流式细胞分析。
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| Isolation and in vivo Transfer of Antigen Presenting Cells
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Author:
Date:
2014-10-05
[Abstract] Transfer of antigen presenting cells in vivo is a method used by immunologists to examine the potency of antigen presentation by a selected population of cells. This method is most commonly used to analyze presentation of protein antigens to MHC class I or II restricted T cells, but it can also be used for studies of nonconventional antigens such as CD1-presented lipids. In a recent study focusing on CD1d-restricted glycolipid antigen presentation to Natural Killer T cells, we compared antigen presenting properties of splenic B cells, CD8αPos dendritc cells (DCs) and CD8αNeg DCs (Arora et al., 2014). This protocol describes the detailed method used for isolation of these cell populations, and their transfer into recipient mice to analyze their ...
[摘要] 体内转移抗原呈递细胞是免疫学家用来检查所选择的细胞群体的抗原呈递效力的方法。该方法最常用于分析蛋白质抗原对MHC I类或II类限制性T细胞的呈递，但其也可用于非常规抗原如CD1呈递脂质的研究。在最近关于CD1d限制性糖脂抗原呈递到天然杀伤T细胞的研究中，我们比较脾B细胞，CD8αpos 树突细胞（DC）和CD8α Neg sup> DC（Arora et al。，2014）。该方案描述了用于分离这些细胞群体和将其转移到受体小鼠中以分析其抗原呈递性质的详细方法。作为总单核细胞的百分比，平均脾脏含有约1-3％骨髓树突状细胞（DC）。在绝对数量上，这转换为大约0.6-1.8×10 6个DC。为了增强脾中DC的数目，向小鼠皮下注射来自经过工程改造以表达fms相关的酪氨酸激酶3配体（Flt3L）的培养的黑素瘤细胞系（B16.Flt3L）的细胞（Mach等人。，2000）。该蛋白是与集落刺激因子-1同源的生长因子，并在造血干细胞的分化中起关键作用。将这种蛋白质作为纯化的蛋白质施用到小鼠中导致多个器官中的CD8αpos pos和CD8α阳性DC亚群的扩增。在已经植入过表达这种蛋白质的肿瘤细胞的小鼠中也观察到类似的扩增（Mach等人，2000）。根据我们的经验，来自具有可触知的B16.Flt3L肿瘤的小鼠的脾脏中高达60％的总单核细胞可以是CD11c阳性树突细胞，从而得到高达5×10 ...
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