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首页> 外文期刊>Clinical hemorheology and microcirculation >Applying XTT, WST-1, and WST-8 to human chondrocytes: A comparison of membrane-impermeable tetrazolium salts in 2D and 3D cultures
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Applying XTT, WST-1, and WST-8 to human chondrocytes: A comparison of membrane-impermeable tetrazolium salts in 2D and 3D cultures

机译：应用XTT，WST-1和WST-8对人软骨细胞：2D和3D培养物中膜渗透渗氮碳盐的比较

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Abstract.BACKGROUND: Tetrazolium-based assays are optimized to assess proliferation/toxicity of monolayer or suspension cells in microtiter plates. With regard to tissue engineering and regenerative medicine the need for in vivo like 3D microtissues has an increasing relevance. Applying tetrazolium-based assays to 3D culture systems is technically more challenging. The composed microenvironment may influence the assay standards, e.g. equal distribution of tetrazolium. OBJECTIVE: Evaluation of membrane-impermeable tetrazolium salt-based assays with regard to spheroid culture (3D) of human chondrocytes. METHODS: Chondrocytes were isolated from human articular cartilage. XTT, WST-1, and WST-8 were applied to monolayer cells (2D, varying cell numbers) and spheroids (3D, different sizes) in 96well plates. Formazan formation was measured spectrophotometrically after different incubation periods. Evaluation was done using phase contrast microsopy (toxicity),analyzing the correlation of cell number and absorbance signals (Gompertz function), and document signal over background ratio.
机译：Abstract.Background：基于四唑鎓的测定经过优化，以评估微量滴定板中单层或悬浮细胞的增殖/毒性。关于组织工程和再生药物，在类似3D微发布等体内的需要增加了相关性。将基于四唑基的测定应用于3D培养系统在技术上更具挑战性。组成的微环境可能影响测定标准，例如，等唑的平等分布。目的：对人软骨细胞的球体培养（3D）的膜渗透渗透盐的测定评价。方法：从人关节软骨中分离有软骨细胞。 XTT，WST-1和WST-8施用于单层电池（2D，不同的细胞数）和96个槽板中的球状细胞（3D，不同尺寸）。在不同孵育期后测定甲藻形成。使用相位对比微晶（毒性）进行评估，分析细胞数和吸光度信号（GOMPertz函数）的相关性和通过背景比的文档信号的相关性。

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