用一组细胞测定法测定高血糖诱导的epc功能障碍:实验小鼠和人类模型系统的验证

Kadambari Dixit, M. Kanitkar, Sheetal Kadam, Rucha Deshpande, V. Kale
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引用次数: 2

摘要

尽管人类和小鼠内皮祖细胞(EPCs)通常用于研究,但由于我们对源特异性应激差异反应的理解有限,因此只能对结果进行不完美的分析。尽管常规使用的细胞和功能分析在单源测试系统中对EPC功能障碍(EPD)的检测是有效的,但缺乏一种能够检测高葡萄糖(HG)和/或糖尿病(DM)诱导的EPD的通用检测系统,无论其来源或部位如何。为了弥补这一缺陷,我们比较了两种细胞来源的测试系统。对各种细胞检测的敏感性比较显示,在所有检测中,只有集落形成检测(CFU)在两种检测系统中对糖尿病/高糖的反应相似,而细胞粘附检测(CAA)、增殖潜力和活力对HG的反应不同。另一方面,小管形成、趋化迁移检测以及CXCR4和VEGFR2 mRNA表达等功能检测在体外和体内均受到HG的影响。有趣的是,研究的其他参数,如一氧化氮、活性氧(ROS)和锰超氧化物歧化酶(MnSOD)对HG和DM暴露的反应不同。在此基础上,我们提出了一组检测方法,包括CFU、小管形成、化学迁移和CXCR4和VEGFR2 mRNA表达,可以准确检测HG / dm诱导的EPD,而不考虑各种系统因素。这些检测还将增强数据集的一致性,并提高人类和小鼠系统中EPD检测的准确性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Determination of hyperglycaemia-induced epc dysfunction using a panel of cellular assays: Validation of experimental murine and human model systems
Although human and murine Endothelial Progenitor Cells (EPCs) are routinely used for research, the results can only be imperfectly analyzed due to our limited understanding of source-specific differential responses to stress. Although the routinely used cellular and functional assays are effective for detection of EPC dysfunction (EPD) in single source test systems, there is lack of a universal detection system capable of detecting high glucose (HG) and/or Diabetes mellitus (DM)-induced EPD irrespective of source or site. To remedy this lacuna we compared the test systems from both cell sources. Comparison of sensitivity of various cellular assays revealed that of all the assays performed, only colony formation assays (CFU) showed comparable responses to diabetes/high glucose in both test systems, while cell adhesion assay (CAA), proliferation potential and viability differed in their responses to HG. On the other hand, the functional assays i.e. tubule formation, chemotactic migration assay and CXCR4 and VEGFR2 mRNA expression were uniformly affected by HG in vitro and DM in vivo. Interestingly, other parameters studied i.e. nitric oxide, reactive oxygen species (ROS) and manganese superoxide dismutase (MnSOD) showed dissimilar responses to HG and DM exposure. On this basis, we propose a panel of assays comprising CFU, tubule formation, chemotactic-migration and CXCR4 and VEGFR2 mRNA expression that can accurately detect HG-/DM-induced EPD irrespective of various systemic factors. These assays will also enhance uniformity across data sets and increase accuracy of EPD detection in human and murine systems.
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