Godoy-Silva Ruben, Mollet Mike, Chalmers Jeffrey J
Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W 19th Ave., Columbus, Ohio 43210, USA.
Biotechnol Bioeng. 2009 Mar 1;102(4):1119-30. doi: 10.1002/bit.22146.
The effect of hydrodynamic forces on animal cell cultures, while extensively studied, still lacks significant, fundamental understanding. A previous manuscript reported on the acute exposure of CHO cells to hydrodynamic forces in a second generation convergent-divergent microfluidic device (Mollet et al., 2007). In this study, the use of this device is extended in a proof of concept system in which suspended animal cells, grown in a typical bioreactor, are subjected to chronic exposure of moderately high levels of hydrodynamic forces by way of a continuous recycle loop between the bioreactor and the microfluidic device. A strain of CHO cells (CHO-6E6) was grown in a batch culture under controlled pH, temperature, and dissolved oxygen conditions. At mid exponential stage of growth in the bioreactor the recycle flow was initiated. The cells either stopped growing or started dying at EDR values that were significantly lower (one to two orders of magnitude) than those previously reported to kill cells from a single, acute exposure. These observations allow further refinement in the design of bioprocess equipment since it provides a more accurate threshold, above which one does not want to subject animal cells to continuous exposure to specific levels of hydrodynamic forces.
尽管对流体动力对动物细胞培养的影响已进行了广泛研究，但仍缺乏重要的、基础性的认识。之前的一篇论文报道了在第二代收敛-发散微流控装置中CHO细胞对流体动力的急性暴露情况（莫莱等人，2007年）。在本研究中，该装置在一个概念验证系统中的应用得到了扩展，在这个系统中，在典型生物反应器中生长的悬浮动物细胞，通过生物反应器和微流控装置之间的连续循环回路，受到中等高水平流体动力的长期暴露。一株CHO细胞（CHO-6E6）在控制pH值、温度和溶解氧条件下的分批培养中生长。在生物反应器生长的指数中期阶段开始循环流动。细胞在比之前报道的单次急性暴露杀死细胞的EDR值显著更低（低一到两个数量级）时，要么停止生长，要么开始死亡。这些观察结果有助于进一步优化生物工艺设备的设计，因为它提供了一个更准确的阈值，超过这个阈值就不希望动物细胞持续暴露于特定水平的流体动力。
