Laboratoire des Sciences du Génie Chimique, CNRS-ENSIC-ENSAIA, Institut National Polytechnique de Lorraine, BP 451, 54001 Nancy Cedex, France.
Biotechnol Bioeng. 1990 Sep;36(6):623-9. doi: 10.1002/bit.260360610.
The production of prourokinase (PUK) by a human kidney tumor cell line is studied in long term cultures. Cells are grown on microcarriers which are retained inside the reactor by sedimentation or with a spin filter. Two modes of operation are compared: feed harvest at an average medium exchange rate of 0.3 d(-1) and continuous perfusion at a higher dilution rate of 1.5 d(-1). In the two systems a stable production of PUK has been maintained for more than 400 h. Kinetics of cellular growth, nutrient consumption, and metabolite and PUK excretion are similar. After an initial rapid growth period, one observes a 10-fold reduction in all the cellular metabolic activities during the stationary phase. Continuous perfusion yields a higher cell density (7 x 10(6) cells.mL(-1)) than feed harvest (3 x 10(6) cells.mL(-1)), which results in a twofold increase in the reactor productivity. But higher final enzyme activities, about 250 ru.mL(-1), are obtained in the feed harvest recovered medium than in the perfusion medium, 100-150 ru.mL(-1). The cumulative medium consumption per mass of product is the same in the repeated batch and in the continuous operation mode.
人肾肿瘤细胞系产生尿激酶原（PUK）的长期培养研究。细胞在微载体上生长，通过沉降或旋转过滤器保留在反应器内。比较两种操作模式：以平均介质交换率 0.3 d(-1)的分批收获和以更高稀释率 1.5 d(-1)的连续灌注。在这两种系统中，PUK 的稳定生产已经维持了 400 多小时。细胞生长、营养物质消耗以及代谢物和 PUK 排泄的动力学相似。在初始快速生长阶段之后，在静止期观察到所有细胞代谢活性降低 10 倍。连续灌注产生的细胞密度（7 x 10(6)个细胞.mL(-1))高于分批收获（3 x 10(6)个细胞.mL(-1))，这导致反应器生产率提高了两倍。但是在分批收获回收培养基中获得的最终酶活性较高，约 250 ru.mL(-1)，而在灌注培养基中约为 100-150 ru.mL(-1)。在重复批次和连续操作模式中，每单位产品的累积介质消耗相同。
