HEKPlus SFM should be supplemented with L-alanyl-glutamine or L-glutamine prior to use. The medium contains 0.1% Poloxamer 188, a surfactant shown to enhance cell yield and reduce cell aggregation. HEKPlus SFM also includes human transferrin and recombinant human insulin that have been shown to improve cell proliferation.
This medium has been optimized for use with ATCC HEK293T/17 SF suspension cells (ATCC ACS-4500) and GeneX Plus transfection reagent (ATCC ACS-4004) to reproducibly express a wide range of proteins at high levels. HEK293T/17 SF cells are pre-adapted to the medium and can be cultured in suspension and directly transfected in HEKPlus SFM using GeneXPlus transfection reagent. Transfection complexes can be prepared in the HEKPlus medium as well. In general, HEK293 cells currently adapted to serum-free medium can be subcultured directly using higher seeding densities. For cells currently adapted to serum containing medium, sequential adaptation to HEKPlus SFM is recommended.