• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biotechnology Progress >Investigation of Expression and Activity Levels of Primary Rat Hepatocyte Detoxication Genes Under Various Flow Rates and Cell Densities in Microfluidic Biochips
【24h】

Investigation of Expression and Activity Levels of Primary Rat Hepatocyte Detoxication Genes Under Various Flow Rates and Cell Densities in Microfluidic Biochips

机译：不同流率和细胞密度下微流生物芯片中大鼠原代肝细胞解毒基因表达和活性水平的研究

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

We investigated the behavior of primary rat hepatocytes in biochips using a microfluidic platform (the integrated dynamic cell culture microchip). We studied the effects of cell inoculation densities (0.2-0.5 X 10~6 cellslbiochip) and peifusion flow rates (10, 25, and 40 μL/ min) during 72 h of perfusion. No effects were-observed on hepatocyte morphology, but the levels ofmRNA and CYP1A2 activity were found to be dependent on the initial cell densities and flow rates. The dataset made it possible to extract a best estimated range of parameters in which the rat hepatocytes appeared the most functional in the biochips. Namely, at 0.25 X 106 inoculated cells cultivated at 25 μLlmin for 72 h, we demonstrated better induction of the expression of all the genes analyzed in comparison with other cell densities and flow rates. More precisely, when primary rat hepatocytes were cultivated at these conditions, the time-lapse analysis demonstrated an over expression of CYP3A1, CYP2B1, ABCClb and ABCC2 in the biochips when compared to the postextraction levels. Furthermore, the AHR, CYP1A2, GSTA2, SULT1A1, and UGT1A6 levels remained higher than 50% of the postextraction values whereas values of HNF4a, CEBP, and PXR remained higher than 20% during the duration of the culture process. Nevertheless, an important reduction in mRNA levels was found for the xenosensors CAR and FXR, and the related CYP (CYP2E1, CYP7A1, CYP3A2, and CYP2D2). CYP1A2 functionality was illustrated by 700 ± 100 pmol/h/10~6 cells resorufin production. This study highlighted the functionality in optimized conditions of primary rat hepatocytes in parallelized microfluidic cultures and their potential for drug screening applications.
机译：我们使用微流控平台（集成动态细胞培养微芯片）调查了生物芯片中原代大鼠肝细胞的行为。我们研究了在72小时的灌注过程中细胞接种密度（0.2-0.5 X 10〜6 cellslbiochip）和灌注流速（10、25和40μL/ min）的影响。没有观察到对肝细胞形态的影响，但发现mRNA和CYP1A2活性的水平取决于初始细胞密度和流速。该数据集使得提取最佳估计范围的参数成为可能，其中大鼠肝细胞在生物芯片中表现出最大的功能。即，在0.25 X 106接种的细胞中，在25μLlmin下培养72小时，与其他细胞密度和流速相比，我们证明了对所有分析基因表达的更好诱导。更准确地说，当在这些条件下培养原代大鼠肝细胞时，延时分析表明，与提取后的水平相比，生物芯片中的CYP3A1，CYP2B1，ABCC1b和ABCC2过表达。此外，在培养过程中，AHR，CYP1A2，GSTA2，SULT1A1和UGT1A6的水平仍高于提取后值的50％，而HNF4a，CEBP和PXR的值仍高于20％。但是，发现异种传感器CAR和FXR以及相关的CYP（CYP2E1，CYP7A1，CYP3A2和CYP2D2）的mRNA水平显着降低。 CYP1A2的功能可以通过700±100 pmol / h / 10〜6个细胞的试卤灵产生来说明。这项研究强调了平行微流体培养中原代大鼠肝细胞在最佳条件下的功能性及其在药物筛选应用中的潜力。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号