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采用扫描介电测量法对感染的 Sf-9 昆虫细胞培养物进行在线监测,并与离线生物量测量进行比较。

On-line monitoring of infected Sf-9 insect cell cultures by scanning permittivity measurements and comparison with off-line biovolume measurements.

机构信息

Pharmaceuticals Division, F. Hoffmann-La Roche AG, Pharma Research Basel, Protein Sciences, Basel, 4070, Switzerland,

出版信息

Cytotechnology. 2007 Dec;55(2-3):115-24. doi: 10.1007/s10616-007-9093-0. Epub 2007 Oct 11.

Abstract

Two infected Sf-9 cell cultures were monitored on-line by multi-frequency permittivity measurements using the Fogale BIOMASS SYSTEM((R)) and by applying different off-line methods (CASY((R))1, Vi-CELLtrade mark, packed cell volume) to measure the biovolume and the mean diameter of the cell population. During the growth phase and the early infection phase the measured permittivity at the working frequency correlated well with the different off-line methods for the biovolume. We found a value of 0.67 pF cm(-1) permittivity per unit of total biovolume (CASY) (muL mL(-1)). After the maximum value in the permittivity was reached, i.e. when the viability of the cultures decreased significantly, we observed different time courses for the biovolume depending on the applied method. The differences were compared and could be explained by the underlying measurement principles. Furthermore, the characteristic frequency (f(C)) was calculated from the on-line scanning permittivity measurements. The f(C) may provide an indication of changes in cell diameter and membrane properties especially after infection and could also be an indicator for the onset of the virus production phase. The changes in f(C) were qualitatively explained by the underlying equation that is correlating f(C) and the properties of the cell population (cell diameter, intracellular conductivity and capacitance per membrane area).

摘要

用 Fogale BIOMASS SYSTEM((R)) 进行多频介电测量对 2 个感染 Sf-9 细胞培养物进行在线监测,并应用不同的离线方法(CASY((R))1、Vi-CELL 商标、细胞体积)测量细胞群体的生物体积和平均直径。在生长阶段和早期感染阶段,工作频率下测量的介电常数与生物体积的不同离线方法密切相关。我们发现每单位总生物体积(CASY)(µL mL(-1)) 的介电常数为 0.67 pF cm(-1)。在介电常数达到最大值后,即当培养物的活力显著下降时,我们观察到不同的生物体积时间过程取决于应用的方法。对差异进行了比较,并可以通过基础测量原理进行解释。此外,还从在线扫描介电测量中计算出特征频率 (f(C))。f(C) 可能是细胞直径和膜特性变化的指示,特别是在感染后,也可能是病毒产生阶段开始的指标。f(C) 的变化通过将 f(C) 与细胞群体的特性(细胞直径、细胞内电导率和单位膜面积的电容)相关联的基本方程进行定性解释。

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