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首页> 外文期刊>Biotechnology Progress >Perfusion Enhanced Polydimethylsiloxane Based Scaffold Cell Culturing System for Multi-Well Drug Screening Platform
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Perfusion Enhanced Polydimethylsiloxane Based Scaffold Cell Culturing System for Multi-Well Drug Screening Platform

机译：用于多孔药物筛选平台的灌注增强型聚二甲基硅氧烷基支架细胞培养系统

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Conventional two-dimensional cultures in monolayer and sandwich configuration have been used as a model for in vitro drug testing. However, these culture configurations do not present the actual in vivo liver cytoarchitecture for the hepatocytes cultures and thus they may compromise the cells liver-specific functions and their cuboidal morphology over longer term culture. In this study, we present a three-dimensional polydimethylsiloxane (PDMS) scaffold with interconnected spherical macropores for the culturing of rat liver cells (hepatocytes). The scaffolds were integrated into our perfusion enhanced bioreactor to improve the nutrients and gas supply for cell cultures. The liver-specific functions of the cell culture were assessed by their albumin and urea production, and the changes in the cell morphology were tracked by immunofluorescence staining over 9 days of culture period. N-Acetyl-Para-Amino-Phenol (acetaminophen) was used as drug model to investigate the response of cells to drug in our scaffold-bioreactor system. Our experimental results revealed that the perfusion enhanced PDMS-based scaffold system provides a more conducive microenvironment with better cell-to-cell contacts among the hepatocytes that maintains the culture specific enzymatic functions and their cuboidal morphology during the culturing period. The numerical simulation results further showed improved oxygen distribution within the culturing chamber with the scaffold providing an additional function of shielding the cell cultures from the potentially detrimental fluid induced shear stresses. In conclusion, this study could serve a crucial role as a platform for future preclinical hepatotoxicity testing.
机译：单层和三明治结构的常规二维培养已用作体外药物测试的模型。但是，这些培养物的配置并不代表肝细胞培养物的实际体内肝细胞结构，因此，它们可能会在长期培养中损害细胞的肝特异性功能及其长方体形态。在这项研究中，我们提出了具有相互连接的球形大孔的三维聚二甲基硅氧烷（PDMS）支架，用于培养大鼠肝细胞（肝细胞）。将支架整合到我们的灌注增强生物反应器中，以改善细胞培养所需的营养和气体供应。通过培养细胞的白蛋白和尿素产量评估其肝特异性功能，并在培养9天的时间内通过免疫荧光染色跟踪其细胞形态的变化。 N-乙酰基-对-氨基-苯酚（对乙酰氨基酚）被用作药物模型来研究细胞在我们的支架生物反应器系统中对药物的反应。我们的实验结果表明，灌注增强的基于PDMS的支架系统提供了更有利的微环境，在肝细胞之间具有更好的细胞间接触，并在培养期间保持了培养特定的酶功能及其长方体形态。数值模拟结果进一步表明，利用支架改善了培养室内氧气的分布，提供了额外的功能，可保护细胞培养物免受可能有害的流体诱导的剪切应力的影响。总之，这项研究可以作为未来临床前肝毒性测试的平台发挥关键作用。

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