novo nordisk pharmatech employee in the laboratory

Case study HEK293 cells

We have explored the effects of recombinant insulin on viral vectors production in HEK293 cells. The results are presented in a case story, and you can read the conclusion here:

The background for the study:

Viral systems are commonly used for the delivery of transgenes for cell & gene therapies and vaccine production. The expansion of this area creates increasing needs for production scale-up, optimized processes, and increased yields. For safety reasons, animal component-free media, like chemically defined (CD) cell culture media with added metabolites, is widely adopted for virus production. In this study, we investigated the effect of recombinant insulin as a supplement in CD media on adeno-associated virus (AAV) and lentivirus (LV) production, using HEK 293 cell line. Viral production was measured at different concentrations (5, 10, and 20 mg/L) of recombinant insulin, and at different addition times (during transfection, 2 hours before transfection, or 4 hours after transfection).

The description of the study:

Three different AAV serotypes were studied (AAV-2, AAV-5, and AAV-8). A three-plasmid system was transferred into HEK293 cells. After 72 hours, AAV particles were collected, the content of ITR gene was detected by qPCR, and the titre (VG/mL) of AAV was calculated. For LV production, a four-plasmid system containing GFP tags was transferred into HEK293 cells. 72 hours post transfection, GFP expression was measured using FACS and the titre (Tu/mL) of LV was calculated.

 

If you wish to read the case study, you can download it and a scientific poster here.