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Ex vivo Non-invasive Assessment of Cell Viability and Proliferation in Bio-engineered Whole Organ Constructs

机译：生物工程全器官构建物中细胞活力和增殖的离体非侵入性评估

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Decellularized organ scaffolds allow whole organ regeneration and study of cell behavior in three-dimensional culture conditions. Cell viability within the bio-engineered organ constructs is an essential parameter reflecting the performance of participating cells during long-term ex vivo culture, and is a prerequisite for further functional performance. Resazurin-based redox metabolic assays have been used to monitor cell viability in both two- and three-dimensional cell cultures. Here we developed a method for monitoring cell viability and proliferation in bio-engineered organ constructs using a resazurin perfusion assay. This method allows non-invasive, repetitive and rapid estimation of viable cell numbers during long-term ex vivo culture. As a proof-of-principle, we assessed the performance of two different endothelial sources and the impact of different perfusion programs on endothelial viability after re-endothelialization of decellularized lung scaffolds. The resazurin-based perfusion assay revealed changes in endothelial viability and proliferation during long-term ex vivo culture, which was consistent with histological assessment at different time points. Finally, we showed that this method could be used for assessment of proliferation and cytotoxicity after pharmacological treatment on a three-dimensional non-small cell lung cancer culture model.
机译：去细胞的器官支架可以在三维培养条件下使整个器官再生并研究细胞行为。生物工程器官构建体内的细胞生存力是反映长期体外培养过程中参与细胞性能的重要参数，并且是进一步发挥功能的先决条件。基于刃天青的氧化还原代谢测定法已用于监测二维和三维细胞培养物中的细胞活力。在这里，我们开发了一种利用刃天青灌注测定法监测生物工程器官构建物中细胞活力和增殖的方法。这种方法可以在长期离体培养过程中以非侵入性，重复性和快速的方式估计活细胞数。作为原理的证明，我们评估了脱细胞肺支架重新内皮化后两种不同内皮来源的性能以及不同灌注程序对内皮活力的影响。基于刃天青的灌注测定揭示了长期离体培养期间内皮细胞活力和增殖的变化，这与在不同时间点的组织学评估一致。最后，我们表明该方法可用于在三维非小细胞肺癌培养模型上进行药理治疗后评估增殖和细胞毒性。

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