CN2006型生物人工支持装置恒温培养系统对胎肝细胞活性的影响

The Effect of CN2006 Biological Artificial Liver Support Device for Cultivating Box with Constant Temperature Control System on Cell Activity of Fetal Liver Cells

目的观察细胞活性,客观评价CN2006型生物人工肝支持装置恒温培养箱控制系统,为生物人工肝的临床应用提供依据。方法选择16周正常引产的人胎肝,胎肝组织采用两步灌流法进行消化分离。在培养条件为37℃、5%CO2、湿度100%下培养24 h后,倒置相差显微镜观察细胞的生长及形态变化;用MTT法检测细胞活力,以2×105/ml细胞数在酶标仪上所测吸光度作为对照;同时,检测胎肝细胞清除氨的能力,设无胎肝细胞的反应器为空白对照。结果胎肝细胞培养24 h后,在倒置相差显微镜下观察发现细胞呈集落状生长。在装置内反应器中培养24 h后,胎肝细胞在装置的反应器中生长良好,而且得到了增殖。细胞在装置内反应器中培养24 h后,加入氯化铵溶液,通过在不同时间检测氨的浓度表明在装置内反应器中培养的肝细胞具有生物转化功能。结论细胞在CN2006型生物人工肝支持装置中不仅生长良好,而且还具有生物转化功能,说明设计的恒温培养系统适宜细胞生长环境,为生物人工肝的临床治疗提供了坚实的理论基础。

O bjective This article aims to observe the cell activity and objectively evaluate CN2006 biological artificial liver support device for cultivating box with constant temperature control system, providing the basis for clinical application of bioartificial liver. Methods To choose human fetal liver of normal induction with 16 weeks. Fetal liver tissue was digested and separated using the two-step perfusion method. After the cultivation for 24 h in the culture conditions of 37℃, 5%CO2, 100%humidity, observe the cell growth and morphological changes under the inverted phase-contrast microscope. Cell viability was detected by MTT, taking 2× 105/ml cell number on the enzyme labelling instrument measured absorbance as a control. Meanwhile, detect the ability of fetal liver cells in removing ammonia, and set a reactor of free fetal liver cells as blank control. Results After culturing the fetal liver cells for 24 h, cells showed the colony growth observed in the inverted phase contrast microscope. After culturing for 24 h in the device inside the reactor, fetal liver cells grew well, and got the proliferation;Adding ammonium chloride solution, it showed cultured liver cells in the device inside the reactor with functions of biological transformation by detecting the concentration of ammonia at different time. Conclusion Cells in CN2006 biological artificial liver support device do not only grow well, but also have the function of biological transformation, which shows that the culturing system with constant temperature is suitable for cell growth, and provides a solid theoretical basis for the clinical treatment of biological artificial liver.