Gene therapy is a promising approach for treating various genetic disorders and diseases. A critical component of gene therapy is the production of viral vectors, which serve as delivery vehicles for therapeutic genes. Human Embryonic Kidney 293 (HEK293) cells have become a widely used platform for viral vector production due to their efficiency in transfection and ability to support viral replication. However, optimizing HEK293 cell cultures for large-scale production of viral vectors remains a challenge in making gene therapies more accessible and cost-effective. This article summarizes key concepts discussed in a webinar by Rui de Paula, Scientist and Instructor at the Jefferson Institute for Bioprocessing (JIB), and Mike Lesch, Regional Sales…