Human embryonic kidney cells, which are naturally adherent, find wide use in the production of viral vectors for several reasons in addition to their high transfection level and rapid growth rate, necessary for the expression of adeno-associated viral (AAV) vectors. Expression of the SV40 large T-antigen in the HEK293-T cell line, meanwhile, supports increased production of AAV, retroviral and lentiviral vectors.

The first gene therapies to reach the market were developed based on viral vectors produced via adherent cell culture and are commercially produced using this technology. Doing so is possible because the patient populations for these products are quite small and the batch sizes needed are amenable to production in flatware such as Corning’s HYPERStacks® and Thermo Fisher’s Nunc™ Cell Factory™ systems. Even today, adherent platforms may be suitable for production of lentiviral vectors used to produce gene-modified cell therapies and gene therapies based on certain AAV serotypes that require smaller volumes. As improvements and technology evolves, fixedbed bioreactors are also enabling more efficient and cost-effective larger-scale manufacturing via adherent cell culture.

Demand for AAV vectors is expected to increase dramatically, however, as the significant numbers of candidates in the clinic, many of which target more prevalent diseases with larger patient populations that must receive bigger and perhaps multiple doses, move closer to commercialization. To meet this greater demand, many gene therapy developers are shifting toward suspension-based platform manufacturing processes for AAV vectors. Scalable suspension cell culture using adapted HEK293 cells in chemically defined media is considered more practical for producing larger batches, and in fact some existing viral-vector manufacturers have indicated they will be transitioning production from adherent to suspension.

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