Evolution Of An Adherent HEK293 Cell Line Into A cGMP Serum-free Suspension Cell Line For Universal AAV, LVV, And Ad Vector Production

HEK293 cells, originally derived from human embryonic kidney cells, are extensively used in viral vector production due to their versatility, which includes a high transfection efficiency and rapid growth. For adeno-associated virus (AAV) production, HEK293 cells are particularly effective because the constitutive expression of the adenovirus early protein E1A acts as a transactivator for the AAV P5 promoter. This makes HEK293 cells an ideal choice for producing recombinant AAV (rAAV) when co-transfected with helper plasmids, AAV Rep/Cap, and ITR-GOI (gene of interest) plasmids.

Early gene therapies relied on viral vectors produced through adherent cell culture, a suitable method for treatments targeting smaller patient groups. Systems such as Corning’s HYPERStacks® and Thermo Fisher’s Nunc™ Cell Factory™ allowed for this type of production. Adherent platforms remain useful for generating lentiviral vectors used in gene-modified cell therapies and certain AAV serotypes where high yields are not essential. However, advances like fixed-bed bioreactors have improved the efficiency of large-scale production using adherent methods, though they come with higher costs.

With growing demand for AAV vectors, driven largely by ongoing clinical trials targeting more prevalent diseases with larger patient populations, the need for expanded manufacturing capacity has increased. Scalable production platforms capable of delivering high-quality viral products are essential. Suspension cell culture, using HEK293 cells adapted to grow in chemically defined, serum-free, animal-product-free media, offers a scalable solution for meeting clinical and commercial demands. As a result, many forward-thinking viral vector manufacturers are shifting from adherent to suspension-based production methods.

Delve into the challenges of scaling viral vector production for cell and gene therapies, and explore innovations in cell line development for AAV, lentivirus (LV), and adenovirus (Ad) production.