293-Hektor

Human Embryo Kidney, serum-free

The human embryo kidney cell line 293 (Sigma Catalogue number. 85120602) adapted to grow in the chemically defined protein- and peptide-free Hektor^™ G medium. Cells grow in suspension and tend to form aggregates in static culture. The cells grow as finely dispersed single cell suspension under optimal agitated culture conditions.

Hektor^™ G medium + 4mM Glutamine + 5 ug/ml Insulin. The exogeneous insulin supplementation can be reduced or totally eliminated under optimal culture conditions. When freezing cells down use culture medium (50:50 conditioned medium:fresh medium) + 10% DMSO.

Viability may be poor on resuscitation and may initially decrease further. Full recovery may take up to 2 weeks. A centrifugation step to remove the cryoprotectant is essential. Rapidly thaw the frozen ampoule in a water bath at 37°C for 1-2 minutes. Transfer the contents to a centrifuge tube and slowly add 5-10ml of pre-warmed growth media. Remove a sample for counting. Centrifuge at 100g for 2-3 minutes to pellet cells and seed at a relatively high density of 5-7 x 10⁵ cells/ml. Leave culture flask upright and observe regularly until viable proliferating cell clumps form. Once established use a split ratio of 1:2 approximately every 4 to 5 days; 5% CO₂; 37°C.

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