HEK293 Cells Cell Culture Information Transfection Information Get HEK293 Transfection Reagent

HEK293 Cell Line

human embryonic kidney cells

Cell Subculture Protocol

HEK293 are rounded cells that grow in suspension in cell culture (although, initially they were an adherent cell line). HEK cells should be grown in a complete SFMII growth medium supplemented with 4mM L-glutamine. Cells should be incubated at 37.0°C in 5% CO2 and expected to double approximately every 34 hours.

Subculture Protocol for HEK293:

Renew medium 2-3 times a week, and keep cell count between 1 x 10^5 and 3 x 10^5 cells/mL. Do no allow cell concentration to exceed 1 x 10 ^ 6 cells/mL

Cells can also be subcultured by centrifuging and resuspending in fresh medium:

  1. Transfer suspension to centrifuge tube
  2. Rap side of emptied flask to remove adherent cells
  3. Resuspend newly dislodged cells in 5 mL medium and aspirate with a small pipette to disperse clumps
  4. Pour resuspended cells into centrifuge tube with rest of suspension
  5. Centrifuge at 125xg for 5-7 minutes
  6. Remove supernatant and resuspend cells with fresh medium

To preserve:

Freeze cells in conditioned growth medium supplemented with 10% (v/v) DMSO. Store in the liquid nitrogen vapor phase or at -135 °C.

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HEK293 (ATCC)

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