生物人工肝支持系统生物反应器的建立及体外转流试验

目的 探讨由微载体培养的L－02人肝细胞和中空纤维舱构成的生物反应器的生物效能。方法 采用微载体培养高浓度L－02人肝细胞，同时使用中空纤维型生物反应器和血泵等共同构成生物人工肝系统。在体外转流试验中观察循环液中游离胆红素、葡萄糖、白蛋白、谷草转氨酶浓度的变化以及实验对肝细胞的影响等。结果 L－02肝细胞能很好地粘附于cytodex 3微载体上形成微载体诱导的肝细胞聚集体；移入生物反应器内进行体外循环，4h后可见循环液中游离胆红素和葡萄糖浓度显著降低，分别为27.1μmmol/L和1.75mmol/L；白蛋白含量明显增加，为15．21mg/L；肝细胞仍有较高活力达75％。结论 本实验所建立的生物反应器作为生物人工肝系统的核心组件具备一定的生物合成和解毒代谢功能，为进一步建立混合型生物人工肝支持系统奠定了基础。

Establishment and preliminary applications of bioreactor in bioartificial liver system

ObjectiveTo investigate the biological function of bioreactor consisting of cultivated human liver cell line L-02 on microcarriers and hollow fiber tube. MethodsHigh concentration L-02 human hepatocytes were cultured on microcarriers. Meanwhile, hollow fiber tube was used as a bioreactor and the blood pump employed as circulation driver, which was composed of simple BAL system and tried in experiment. The changes in concentrations of TBIL, ABL, GLU and AST were observed under the condition of in vitro circulation. ResultsThe L-02 human hepatocytes were perfectly attached to cytodex 3 microcarriers, which called microcarrier-induced liver cell aggregation (MILCA). During the 4 hours of in vitro circulation, marked increase of albumin and decrease of bilirubin and glucose were found. At the end of the in vitro circulation, the values of bilirubin, albumin and glucose were 27.1 umol/L, 15.2 mg/L and 1.75 mmol/L, respectively. The viability of hepatocytes in 75% cells was observed after the experiment. ConclusionsThe bioreactor used as the core of ABL system has functions of biosynthesis and biotransformation. It can become an effective bioartificial liver device.