Chen Antony K, Latz Michael I, Frangos John A
Department of Bioengineering, University of California-San Diego, La Jolla, California, USA.
Biotechnol Bioeng. 2003 Jul 5;83(1):93-103. doi: 10.1002/bit.10647.
Bioluminescent dinoflagellates are flow-sensitive marine organisms that produce light emission almost instantaneously upon stimulation by fluid shear in a shear stress dose-dependent manner. In the present study we tested the hypothesis that monitoring bioluminescence by suspended dinoflagellates can be used as a tool to characterize cellular response to hydrodynamic forces in agitated bioreactors. Specific studies were performed to determine: (1) impeller configurations with minimum cell activation, (2) correlations of cellular response and an integrated shear factor, and (3) the effect of rapid acceleration in agitation. Results indicated that (1) at a volumetric mass transfer coefficient of 3 x 10(-4) s(-1), marine impeller configurations were less stimulatory than Rushton configurations, (2) bioluminescence response and a modified volumetric integrated shear factor had an excellent correlation, and (3) rapid acceleration in agitation was highly stimulatory, suggesting a profound effect of temporal gradients in shear in increasing cell stimulation. By using bioluminescence stimulation as an indicator of agitation-induced cell stimulation and/or damage in microcarrier cultures, the present study allows for the verification of hypotheses and development of novel mechanisms of cell damage in bioreactors.
生物发光甲藻是对流动敏感的海洋生物,在受到流体剪切力刺激时几乎能瞬间产生发光,且发光强度与剪切应力呈剂量依赖关系。在本研究中,我们检验了这样一个假设:通过悬浮的甲藻监测生物发光可作为一种工具,用于表征在搅拌式生物反应器中细胞对流体动力的反应。进行了具体研究以确定:(1)使细胞活化最小的叶轮配置;(2)细胞反应与综合剪切因子的相关性;(3)搅拌快速加速的影响。结果表明:(1)在体积传质系数为3×10⁻⁴ s⁻¹时,船用叶轮配置比Rushton叶轮配置的刺激性小;(2)生物发光反应与修正的体积综合剪切因子具有良好的相关性;(3)搅拌快速加速具有高度刺激性,表明剪切力的时间梯度对增加细胞刺激有深远影响。通过将生物发光刺激作为微载体培养中搅拌诱导的细胞刺激和/或损伤的指标,本研究有助于验证假设并开发生物反应器中细胞损伤的新机制。