Abu-Reesh I, Kargi F
Department of Chemical Engineering, Washington University, St. Louis, Missouri.
Enzyme Microb Technol. 1991 Nov;13(11):913-9. doi: 10.1016/0141-0229(91)90108-m.
Effects of long-term hydrodynamic shear on hybridoma cells were investigated in a 250-ml continuous stirred-tank reactor (CSTR). Cells grown at steady state were subjected to step changes in agitation rates. Cell viability, glucose consumption, and monoclonal antibody (MAb) production were determined at high agitation rates and compared with the control (100 rev min-1). Impeller tip speeds higher than 40 cm s-1 caused a significant drop in cell concentration and respiration activity, and increased lactate dehydrogenase (LDH) release to the culture medium. Also, high agitation speeds caused a decrease in MAb concentration and an increase in specific glucose consumption rate. The effects of dilution rate and serum concentration on the sensitivity of hybridoma cells to hydrodynamic shear were determined. Serum was found to protect the cells against shear damage and had a significant positive effect on hybridoma growth and MAb production. Shear damage on cells in CSTR was approximated to first-order kinetics. The death rate constant increased sharply at impeller tip speeds above 40 cm s-1.
在一个250毫升的连续搅拌釜式反应器（CSTR）中研究了长期流体动力剪切对杂交瘤细胞的影响。将处于稳态生长的细胞置于搅拌速率的阶跃变化中。在高搅拌速率下测定细胞活力、葡萄糖消耗和单克隆抗体（MAb）产量，并与对照（100转/分钟）进行比较。叶轮尖端速度高于40厘米/秒会导致细胞浓度和呼吸活性显著下降，并增加乳酸脱氢酶（LDH）向培养基中的释放。此外，高搅拌速度会导致MAb浓度降低和比葡萄糖消耗速率增加。测定了稀释率和血清浓度对杂交瘤细胞对流体动力剪切敏感性的影响。发现血清可保护细胞免受剪切损伤，并对杂交瘤生长和MAb产生有显著的积极影响。CSTR中细胞的剪切损伤近似为一级动力学。在叶轮尖端速度高于40厘米/秒时，死亡率常数急剧增加。
