Production of useful biochemicals by immobilized animal cells

为了提高利用动物细胞培养生产有用生物化学物质的效率，开发了一种新的固定化方法。将两株产生抗促红细胞生成素和抗α-淀粉酶单抗的杂交瘤细胞系包埋在海藻酸盐凝胶中，然后用氨基甲酸酯聚合物包被固定化的动物细胞凝胶。通过这种固定化方法，获得了超过107个细胞/毫升凝胶，抗体的体积产率比常规培养提高了近10倍。为了培养锚定依赖细胞，使用了新型大孔纤维素纤维(SIC)。将转化的人内皮细胞固定在该载体上，在沸腾床反应器中进行培养。通过优化培养基及供氧条件，可获得浓度大于10个/ml的碳化硅细胞。通过在碳化硅表面引入带电基团，杂交瘤细胞也被固定化，细胞浓度接近108个/毫升，获得了碳化硅。因此，我们比较了几种方法，发现使用全氟化合物可以提供足够的氧气，以满足高浓度细胞生长的需要，超过107cell/ml。

In order to improved the production of useful biochemicals by animal cell culture, a novel immobilization methods have been developed. Two hybridoma cell lines that produced anti- erythropoietin and anti-alpha -amylase monocional antibody were entrapped in alginate gel and then the immobilized animal cell gel was covered with urethane polymer. By this method of immobilization, more than 10^7 cells/ml gel was obtained and the volumetric productivity of the antibody increased almost 10 times comparing to the regular cultures.In order to culture anchorage depended cells, a novel macroporous cellulose fiber(SIC) was used. Transformed human endothelial cells were immobilized on this support and cultured in a fluidized bed reactor. By optimizing medium and oxygen supply, high concentration of cells more than 10 cells/ml. SIC could be obtained. By introducing a charged group on the surface of SIC, hybridoma cell was also immobilized and high concentration of cells near 10^8 cells/ml. SIC was attained.By regular cultivation of animal cells, oxygen supply is the most serious problem. Therefore, we compared several method and found that enough amount of oxygen for the growth of high concentration of cells more than 10^7 cell/ml could be supplied by using perfluorochemicals.