Influenza virus production in the ReadyToProcess WAVE 25 bioreactor system with presterilized microcarriers

We took an established influenza virus production process and adapted it for use with our new gamma irradiated microcarriers. Find out how the results compared with the original process.

Reduce time and complexity

Adherent Vero cells can multiply only when they have a surface to attach to. Microcarriers provide a large surface area and have been shown to be a suitable choice for growing adherent cells in Cellbag bioreactors.

In the legacy process, Cytodex microcarriers had been successfully used. However, they must be washed, sterilized by autoclaving, and equilibrated before use. These steps add significant time and complexity to the process, especially at scale.

By contrast, presterilized Cytodex Gamma microcarriers can be added straight into the bioreactor with no preparatory steps, so time and complexity is reduced. Another benefit is a lower contamination risk, as Cytodex Gamma can be added to the bioreactor via an aseptic connector in a completely closed process.

 29209415AC_Figure-01-updated

 

The two microcarriers were used side by side in two 10 L Cellbag bioreactors with the same 4 L culture volume. Parameters such as temperature and CO2 levels were maintained at the same levels in both Cellbag bioreactors. The cultures were sampled every 24 hours to check cell density and cell morphology.

The results

The end results show that Cytodex and Cytodex Gamma are comparable in supporting both cell growth and virus production in a ReadyToProcess WAVE 25 rocking bioreactor system. This study demonstrates that using presterilized Cytodex Gamma to reduce time and simplify processing does not compromise on high-quality output.

 

29209415AC_Figure_03

 

Viable cell density in cultures using either Cytodex 3 Gamma or Cytodex 3 microcarriers in 4 L culture volume. Cells were cultured in ReadyToProcess WAVE 25 using a rocking motion setting of either 30% or 100%.

 

TCID50 analysis results

Sample
TCID50
Cytodex 3, 100% rocking motion
107.1/mL
Cytodex 3 Gamma, 100% rocking motion  107.5/mL
Cytodex 3, 30% rocking motion
107.5/mL
Cytodex 3 Gamma, 30% rocking motion
107.7/mL