在添加醋酸钾的营养富集培养基中培养杂交瘤细胞(英)

营养富集、添加醋酸钾和灌注培养都是改善杂交瘤细胞培养的策略在Cp9B细胞的静止分批培养中，添加氨基酸和维生素延长了培养时间，但对最大细胞密度影响不大，在反应器分批培养中，营养物明显促进细胞增殖，最大细胞密度比原来提高35倍．利用添加1g／L醋酸钾的反应器灌注培养实验说明，从普通培养基转到营养富集培养基，反应器中的平均细胞密度、抗体浓度和抗体滴度得到了进一步的提高，在培养上清中，乳酸、氨和丙氨酸大量积累，葡萄糖和某些氨基酸大量消耗，以致于可能造成对细胞生长和代谢的抑制。

HYBRIDOMA CELL CULTURE IN A NUTRIENT-FORTIFIED MEDIUM SUPPLEMENTED WITH POTASSIUM ACETATE

Nutrient fortification, supplementation of potassium acetate (KAc) and cell cycling in perfusionculture were some of the strategies used to improve yields in hybridoma cultures. In static batch culture of Cp9Bcells, fortifications of amino acids and vitamins prolonged the culture span, but could do little in increasingmaximal cell density. The nutrients obviously promoted cell proliferation in bioreactor batch cultures and maximai cell density was 3 .5 times of that in unfortified bioreactor batch culture and 2 .7 times of that in fortifiedstatic batch culture. Comparative experiments of nutrient fortification with additional supplementation of 1 g/LKAc in perfusion cultures in a bioreactor coupled with a cell retention system were Performed. T'be results ofthese experiments revealed that when foe perfusion culture was transferred from unfortified medium to fortified medium, average cell density, monoclonal antibody (MAb) concentration and titer in bioreactor were further increased. In culture supernatant, a large amount of lactate, ammonia and alanine were accumulated while glucose and some amino acids were depleted to such an extent that they might cause suppression ic cell growth andmetabolism .