Bioprozeß- und Bioverfahrenstechnik, Technische Universität Hamburg-Harburg, Denickestr. 15, 21071, Hamburg, Germany.
Cytotechnology. 1995 Jun;19(2):111-24. doi: 10.1007/BF00749766.
A murine hybridoma cell line producing a monoclonal antibody against penicillin-G-amidase and a murine transfectoma cell line secreting a monovalent chimeric human/mouse Fab-antibody fragment were cultivated in three different media (serum-containing, low protein serum-free, and iron-rich protein-free) in flask cultures, stirred reactors and a fixed bed reactor. In static batch cultures in flasks both cell lines showed similar good growth in all three media.In suspension in a stirred reactor, the hybridoma cell line could be cultivated satisfactory only in serum-containing medium. In low protein serum-free medium, Pluronic F68 had to be added to protect the hybridoma cells against shear stress. But even with this supplement only batch, not chemostat mode was possible. In iron-rich protein-free medium the hybridoma cells grew also in continuous chemostat mode, but the stability of the culture was low. The transfectoma cell line did not grow in stirred reactors in any of the three media.Good results with both cell lines were obtained in fixed bed experiments, where the cells were immobilized in macroporous Siran(®)-carriers. The media, which were optimized in flask cultures, could be used without any further adaptation in the fixed bed reactor. Immobilization improved the stability and reliability of cultures of non-adherent animal cells in serum-free media tremendously compared to suspension cultures in stirred reactors. The volume-specific glucose uptake rate, an, indicator of the activity of the immobilized cells, was similar in all three media. Deviations in the metabolism of immobilized and suspended cells seem to be mainly due to low oxygen concentrations within the macroporous carriers, where the cells are supplied with oxygen only by diffusion.
一株产生针对青霉素 G 酰胺酶的单克隆抗体的鼠杂交瘤细胞系和一株分泌单价嵌合人/鼠 Fab 抗体片段的鼠转染瘤细胞系在三种不同的培养基（含血清、低蛋白无血清和富含铁的无蛋白）中进行了培养，包括在摇瓶培养、搅拌式反应器和固定床反应器中。在摇瓶中的静态分批培养中，两种细胞系在三种培养基中均表现出相似的良好生长。在搅拌式反应器中的悬浮培养中，杂交瘤细胞系仅能在含血清的培养基中得到满意的培养。在低蛋白无血清培养基中，必须添加 Pluronic F68 以保护杂交瘤细胞免受剪切力的影响。但即使添加了这种补充剂，也只能进行分批培养，而不能进行恒化培养模式。在富含铁的无蛋白培养基中，杂交瘤细胞也能以连续恒化培养模式生长，但培养的稳定性较低。转染瘤细胞系在三种培养基中的搅拌式反应器中均无法生长。在固定床实验中，两种细胞系都取得了良好的结果，其中细胞被固定在大孔 Siran（®）载体中。在摇瓶培养中优化的培养基可直接用于固定床反应器，无需进一步调整。与搅拌式反应器中的悬浮培养相比，固定化极大地提高了无血清培养基中非贴壁动物细胞培养的稳定性和可靠性。比耗氧速率 an，作为固定化细胞活性的指标，在三种培养基中相似。固定化和悬浮细胞代谢的偏差主要归因于大孔载体内部的低氧浓度，细胞只能通过扩散获得氧气。
