红细胞携氧促进多层平板型生物反应器中肝细胞氧供

目的 生物人工肝作为肝功能衰竭的一种有效支持手段,近年来得到不断的改进,如何解决反应器中的氧供问题成为目前人们研究的焦点.本研究拟通过在循环培养液中加入红细胞为这一问题提供可能的解决方案.方法 将新鲜分离的原代猪肝细胞接种于笔者自主构建的新型生物反应器内.实验分为两组,对照组在反应器内仅加入RPMI1640培养液直接循环;实验组在培养液中同时加入猪红细胞(2.5×1011/L)进行循环,两组循环液均经过膜肺进行氧合.采用血气分析仪检测反应器中氧耗情况,同时检测反应器中葡萄糖消耗及肝细胞的功能表达.结果 实验组反应器中肝细胞氧耗率为对照组的1.5倍,葡萄糖消耗为对照组的2倍.同时,实验组肝细胞的白蛋白分泌及尿素合成各项功能均明显高于对照组.结论 在培养液中加入红细胞能显著改善反应器中肝细胞的氧供,从而提高反应器中细胞的葡萄糖代谢及各种肝特异功能的表达.该方法简便易行,效果明显,有望成为解决反应器中氧供的有效手段.

Enhanced oxygen delivery to primary hepatocytes within a multi-layer flat plate bioreactor via red blood cells

Objective As an effective means of liver function support for acute liver failure, bioartificial liver has seen great progress in recent years. However, the development of this type of device is currently hindered by limited oxygen transport to cultured hepatocytes. In this study we try to resolve this problem by supplementing the circulating medium of the bioreactor with red blood cells.Methods Freshly isolated primary porcine hepatocytes were inoculated into our newly designed bioreactor and were divided into two groups: RPMI1640 was circulated in the control group and porcine red blood cells were added into the culture medium in the experimental group. The culture media in both groups were oxygenated through extra-corporeal membrane oxygenation, and the oxygen consumption rate (OCR) in the bioreactor was measured by a blood gas analyzer. Liver-specific functions and glucose consumption were also determined. Results The OCR of the experimental group was 1.5 fold that of the control group, and the glucose consumption rate was twice that of the control group. The liver-specific functions of the experimental group were significantly higher than those of the control group in terrns of albumin secretion and urea synthesis. Conclusion Supplementing the circulating medium of the bioreactor with red blood cells can significantly improve the oxygen supply in the bioreactor, thereby enhancing the glucose consumption and liver-specific functions of hepatocytes. This method is convenient and effective, and is expected to be an effective means to resolve the problems of oxygen supply in the bioreactor.