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Hybridoma Cell Culture Application Using a BioFlo 110 Benchtop Bioreactor

(1) The Biomass monitor and sensor used in this experiment are not available in the marketplace as they are still undergoing test runs.

When this equipment is available, we will make an announcement in the Culture Club, BioFlo 110 newsletter.

The Biomass sensor uses the principle that under the effect of an electric field between two electrodes, live cells (which have intact plasma membranes) polarize and behave as small condensers. The value of the resulting capacitance is measured by the second pair of electrodes and correlates exactly to the concentration of biomass in the culture environment.

In the first stage of culture (0-30 hrs), the pH and DO were controlled by surface aeration at a rate of 200 ml/min. After 30 hours of culture, the capacitance value began to decrease to 0.42 [pf/cm] at 45 hour, as shown in Figure 3. At this time perfusion was started at 1 liter per day, which rapidly increased the value to 0.73 [pf/cm] after 90 min. This phenomenon occurred three times (and is indicated in Figure 4 as T1, T2 and T3) during the S1 and S2 stages.We believe that the plasma membrane polarization for each cell changed, depending on the cell culture conditions.

(2) The equation below was used to calculate the linear correlation between viable cell densities (determined by NucleoCounter, Trypan Blue) and capacitance (determined by biomass probe).

X [10 6 cells/ml] = 1.386 X C [pf/cm]

Here, X is viable cell density [10 6 cells/ml] and C is capacitance [pf/cm]



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