| In vitro Assays for the Detection of Calreticulin Exposure, ATP and HMGB1 Release upon Cell Death
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Author:
Date:
2016-12-20
[Abstract] Accumulating evidence is revealing the essential role of immune system in cancer treatment. Certain chemotherapeutic drugs can potently induce the release of ‘cell death associated molecular patterns’ (CDAMPs), which accompanies cancer cell demise. CDAMPs can engage corresponding receptors on immune cells and stimulate immune responses to achieve long-term tumor control (Ma et al., 2013; Ma et al., 2014; Yang et al., 2015). Among reported CDAMPs, calreticulin (CALR), ATP and HMGB1 are well known for their immune-stimulatory effect. Here we describe the assays that we applied to measure cell death and these CDAMPs. Briefly, cell death can be analyzed by co-staining of 4’,6-diamidino-2-phenylindole (DAPI) with 3,3’-Dihexyloxacarbocyanine Iodide [DiOC6(3)] or ...
[摘要] 积累的证据表明免疫系统在癌症治疗中的重要作用。某些化学治疗药物可以有效诱导伴随癌细胞死亡的“细胞死亡相关分子模式”（CDAMPs）的释放。 CDAMP可以与免疫细胞上的相应受体结合并刺激免疫应答以实现长期的肿瘤控制（Ma等，2013; Ma等人，2014; Yang＆ em>等，，2015）。在所报道的CDAMP中，钙网蛋白（CALR），ATP和HMGB1是众所周知的免疫刺激作用。在这里我们描述我们应用于测量细胞死亡和这些CDAMPs的测定。简单地说，细胞死亡可以通过4'，6-二脒基-2-苯基吲哚（DAPI）与3,3'-二氧基氧羰基花青氨基碘[DiOC6（3）]或Annexin V的共染色来分析。通过流式细胞术检测。 ATP和HMGB1释放可以通过发光测定和ELISA测定分别进行定量。
背景 乳酸脱氢酶测定和台盼蓝染色是检测细胞死亡的传统方法。我们在这里描述两种可行和经济的解决方案，以通过流式细胞术（FCM）检测凋亡和坏死细胞死亡。 DAPI标记具有破坏完整性（坏死）的细胞，而膜联蛋白V结合磷脂酰丝氨酸（其在凋亡后被外化）。 DiOC6（3）摄取表明线粒体跨膜电位（MTP），MTP的崩解显示凋亡。 DAPI不需要补偿藻红蛋白（PE，其与膜联蛋白V蛋白结合）或DiOC6（3），因此在这些测定中显示优于碘化丙啶（PI）的优点。 ...
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| Coculture between hMADS and Mouse Adult CM
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Author:
Date:
2014-07-20
[Abstract] Heart failure occurring after acute myocardial infarction (MI) is among the main causes of death in western countries. Cell therapies, particularly those based on mesenchymal stem cells (MSC), represent one of the most promising approaches to repair damaged heart tissues. Several reports have provided evidences that injection of mesenchymal stem cells improved heart function following myocardial infarction (Shake et al., 2002; Zimmet and Hare, 2005; Zeng et al., 2007). Nevertheless, the mechanism(s) by which MSC exert their therapeutic action is far from being understood, and further knowledges in this field are required especially to optimize efficiency of current cardiac cell therapies. To assess the regenerative mechanisms developed by MSC in vitro, we ...
[摘要] 急性心肌梗死（MI）后发生的心力衰竭是西方国家死亡的主要原因。细胞疗法，特别是基于间充质干细胞（MSC）的那些，代表了修复受损心脏组织的最有希望的方法之一。几个报道提供了证据，注射间充质干细胞改善了心肌梗塞后的心脏功能（Shake等人，2002; Zimmet和Hare，2005; Zeng等人， 2007）。然而，MSC发挥其治疗作用的机制远未被理解，并且尤其需要在该领域中的进一步知识以优化当前心脏细胞治疗的效率。为了评估由MSC在体外形成的再生机制，我们开发了上述方法，其预期模拟梗塞心脏的典型微环境。该方法包括处于不良状态的小鼠终末分化心肌细胞与本文用作MSC模型的人多能脂肪来源干细胞（hMADS细胞）之间的物种错配共培养物。
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