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首页> 外文期刊>Applied in vitro toxicology. >A Simple Fluorescent Cell-Type-Specific Labeling Method for Cocultured Cell Flow Cytometry Analysis
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A Simple Fluorescent Cell-Type-Specific Labeling Method for Cocultured Cell Flow Cytometry Analysis

机译：一个简单的细胞类型特异的标记荧光Cocultured细胞流式细胞仪的方法分析

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Introduction: Coculture models have been extensively used for assessing the toxicity of fibers and particles. However, once cell lines are mixed in the same tissue culture well, it is difficult to evaluate differential cell toxicity without using specific cell markers, which might not be compatible with assays requiring living cells such as a particle-induced oxidative stress assay by flow cytometry.Materials and Methods: Human alveolar epithelial cells (A549) were specifically labeled with cell proliferation dye—eFluor? 670—before being mixed with phorbol ester-differentiated Tohoku Hospital Pediatrics-1 cells (used as macrophages). The coculture model allowed the toxicity of crystalline silica DQ-12 and DQ-12-PVNO (particles were coated with polyvinylpyridine-N-oxide, a molecule used to quench particle surface reactivity) to be assessed. Particle-induced oxidative stress was evaluated by flow cytometry using a 2′,7′-dichlorodihydrofluorescein diacetate probe (H2DCFDA).Results: A549 cell treatment with a noncytotoxic concentration of an eFluor 670 probe allowed labeled and unlabeled cells to be differentiated using flow cytometry. Cellular oxidative stress induced by phorbol ester or DQ-12 detected by H2DCFDA was not affected by eFluor 670 probe cell treatment.Conclusions: This study showed that specific labeling of live cells before coculture setup allows assays such as oxidative stress assessment by flow cytometry to be conducted on different live cell types from the same coculture models without the use of cell-type-specific markers. Such assays would be of great value in any kind of multiple cell type model in which the effects of chemicals on a given cell population is sought.
机译：作品简介:Coculture模型广泛用于评估的毒性纤维和粒子。混合在同一组织培养好,吗很难评估微分细胞毒性不使用特定的细胞标记,这可能不兼容化验需要生活如粒子诱导细胞氧化应激通过流式细胞仪测定。人肺泡上皮细胞(A549)特别是标以细胞增殖dye-eFluor吗?ester-differentiated东北医院Pediatrics-1细胞(巨噬细胞)使用。允许晶体硅DQ-12的毒性和DQ-12-PVNO(粒子被涂上一层polyvinylpyridine-N-oxide,一个分子淬火粒子表面反应性)评估。流式细胞仪使用评估2 ', 7 ' -dichlorodihydrofluorescein二醋酸盐调查(H2DCFDA)。noncytotoxic eFluor 670探针的浓度允许标记和未标记细胞使用流式细胞仪分化。氧化应激引起的或佛波醇酯DQ-12 H2DCFDA探测到没有影响eFluor 670探测细胞治疗。研究表明,特定标签的活细胞之前coculture设置允许化验等氧化应激评估通过流式细胞术在不同的活细胞类型不使用相同的coculture模型cell-type-specific标记。伟大的价值在任何类型的多种细胞类型模型上的化学物质的影响给定的细胞群。

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