| Protocol for Murine/Mouse Platelets Isolation and Their Reintroduction in vivo
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Author:
Date:
2017-02-20
[Abstract] Platelets and coagulation have long been known to be essential for metastasis in experimental models. In order to study the interactions between tumor cells, platelets and endothelium, we have adapted methods used in coagulation research for the isolation of platelets and their reintroduction into mice. Anti-coagulated murine blood served as the source for platelets. Platelets were separated from other elements of the whole blood by centrifugation. Here the critical elements are first inhibition of coagulation and second isolation and maintenance of the platelets in the presence of inhibitors of platelet activation. We then used the vital dye PKH26 to fluorescently label the platelets. Infusion of these labelled platelets allows microscopic observation of the introduced platelets. After ...
[摘要] 长期以来，已知血小板和凝血酶在实验模型中对转移至关重要。为了研究肿瘤细胞，血小板和内皮之间的相互作用，我们采用了凝血研究中用于分离血小板及其再引入小鼠的适应方法。抗凝血鼠血液作为血小板的来源。通过离心将血小板与全血的其它元素分离。这里的关键因素是在存在血小板活化抑制剂的情况下首先抑制凝血和血小板的第二次分离和维持。然后用生物染料PKH26荧光标记血小板。这些标记血小板的输注允许显微观察引入的血小板。重新引入后，这些血小板看起来正常起作用，占总血小板的约50％。因为它们是荧光标记的，所以它们很容易被识别。最后，可以使用这些方法通过使用来自基因工程小鼠的血小板来确定改变的基因表达在血小板中的特异性效应。这些方法有助于研究组织培养和鼠模型中血小板和肿瘤细胞之间的相互作用。它们也适用于视频显微镜。在这里，我们提供了我们用于小鼠血小板分离的方法的细节，以及它们的染色用于进一步的显微镜和重新引入小鼠。
背景 已知血小板对于转移是必需的，而且在肿瘤生长期间也起作用，更不用说凝块形成。为了容易识别和追踪血小板，我们开发了荧光标记和再灌注血小板的手段。这使得它们可以容易地在不需要免疫染色的组织中鉴定。使用这些方法，我们显示肿瘤细胞和血小板之间的相互作用在转移早期在肿瘤细胞的存活中起关键作用（Im等人，2004; ...
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| C1q Binding to and Uptake of Apoptotic Lymphocytes by Human Monocyte-derived Macrophages
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Author:
Date:
2013-09-05
[Abstract] To characterize macrophage gene expression profiles during the uptake of autologous apoptotic cells, we developed a unique, more physiologic system using primary human monocyte derived macrophages purified via a nonactivating isolation procedure (and in the absence of contaminating platelets, which can release stimulating signals if activated) and autologous lymphocytes as a source of apoptotic cells. The use of autologous cells as the apoptotic target rather than transformed cell lines avoids antigenic stimulation from “nonself” structures at the HLA level but also from “altered self” signals due to the transformation inherent in cell lines.
[摘要] 为了表征在自体凋亡细胞摄取期间的巨噬细胞基因表达谱，我们开发了一种独特的，更生理的系统，使用通过非活化分离程序纯化的原代人单核细胞衍生的巨噬细胞（和在没有污染的血小板，如果激活可以释放刺激信号 ）和自体淋巴细胞作为凋亡细胞的来源。 使用自体细胞作为凋亡靶而不是转化细胞系避免了来自HLA水平上的"非自身"结构的抗原刺激，而且还由于细胞系中固有的转化而导致的"改变的自身"信号。
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