| Field | Specification |
|---|---|
| Species |
HEK293-F cells are a fast-growing, highly transfectable subline derived from the human embryonic kidney 293 (HEK293) cell line. The 'F' designation indicates that these cells have been adapted for growth in suspension cultures, making them particularly useful for large-scale protein production. The cells grow in a variety of serum-free media, facilitating scalable processes in biotechnological and pharmaceutical applications. HEK293-F cells retain the epithelial-like morphology of the parent HEK293 line and are maintained in suspension without the need for attachment to a solid substrate.
These cells are highly efficient at expressing recombinant proteins and are widely utilized in the production of viral vectors for gene therapy, including adenoviral, lentiviral, and retroviral vectors. Their robust growth in suspension and ease of transfection make them ideal for use in transient transfection protocols, where they can produce high yields of protein within a few days post-transfection. This characteristic is critical for rapid production cycles in research and industrial settings. The adaptability of HEK293-F cells to various growth conditions and their capacity for high-density culture enhance their utility in bioprocessing environments.
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- Receptors expressed: Vitronectin
- Protein expression: CEA negative, p53 positive
- Tumorigenic: In nude mice
- Viruses: Transformed with adenovirus 5 DNA
- adenovirus 5 DNA
- cultureMedium: CD293 (Thermo) + L-Glutamin
- supplements: Supplement the medium with 10% FBS and 1% NEAA
- dissociationReagent: Accutase
- doublingTime: 30 hours
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- seedingDensity: 1 x 104 cells/cm2 will yield in a confluent layer in about 4 days
- fluidRenewal: 2 times per week
- postThawRecovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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