Ghani, K., Cottin, S., Kamen, A., & Caruso, M. (2007). Generation of a high-titer packaging cell line for the production of retroviral vectors in suspension and serum-free media. Gene therapy, 14(24), 1705.
DOI: 10.1038/sj.gt.3303039

The aim of this study was to generate a retrovirus packaging cell line that could be used for the production of large clinical batch vectors. Bicistronic vectors containing an internal ribosomal entry site followed by a selection gene were used to express Moloney murine leukemia gag-pol and amphotropic envelope viral proteins in HEK293 cells. The candidate clone (293GP-A2) that was selected as the packaging cell line could release recombinant green fluorescent protein retroviruses at 4x10(7) infectious viral particles per ml. Similar titers were achieved after these cells were adapted to grow in suspension and serum-free media.

The serum-free media was a low-calcium-SFM derived from H-SFM, and was supplemented with 0.1% (v/v) lipid mixture, 0.1% (v/v) bovine serum albumin and 0.1% Pluronic F-68 as specified in the Materials and Methods,under the heading; Cell culture. H-SFM is a commercial product. The precise formulation is proprietary.