Cell culture-produced adeno-associated viral (AAV) vectors have gained momentum as one of the most effective gene and protein delivery tools and are an integral part of both vaccine production and gene therapy development.  

Yet, in order to produce sufficient cell numbers for a given application and to keep the process reliable and reproducible, process scale-up options as well as accurate process parameter control are necessary. Here, bioreactors can be a powerful platform with their precise control over parameters like temperature, pH, medium agitation, gassing composition and others.  

In this study, single-use bioreactors regulated by Eppendorf bioprocess control systems were utilized to establish an efficient and scalable AAV capsid production platform based on Expi293F suspension cell culture. In order to prepare sufficient cell numbers for inoculating the bioreactor culture, a shake flask seed train was established using a CO2 incubator shaker.

Learning Objectives:

1. Learn about advantages and disadvantages of adherent and suspension cell culture for different applications.

2. Get to know different seed train strategies to prepare a sufficient inoculum for your culture.

3. Learn about process development of AAV production in single-use bioreactors.