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The 293T/17 cell line is an immortalized variant of the HEK293 line, derived from human embryonic kidney cells and extensively utilized in research, particularly in the study and production of retroviral and lentiviral vectors. This cell line has been modified to express the SV40 large T antigen, enhancing its utility in viral vector production. The expression of the SV40 large T antigen is a key feature that allows these cells to replicate plasmids containing the SV40 origin of replication, significantly increasing the yield of plasmid DNA in transient transfection procedures. This feature is particularly beneficial for the production of viral vectors.
293T/17 cells are essential in the production of viral vectors like retroviruses and lentiviruses. They efficiently produce viral particles due to their ability to amplify transfected plasmids and support viral assembly and release. This makes them a vital tool in gene therapy research, where these vectors are used to deliver genetic material into host cells. The cells exhibit high transfection efficiency, crucial for the successful introduction and expression of foreign DNA during vector construction. This high efficiency enables the study of gene function and the generation of recombinant proteins effectively.
The robust capabilities of the 293T/17 cell line make it invaluable for both basic scientific research and therapeutic applications. It is widely used in molecular biology and genetic engineering for protein expression, gene function analysis, and the development of novel gene therapies. The cell line's efficiency in producing viral vectors facilitates experiments requiring genetic material delivery, making it a cornerstone in the field of virology. The 293T/17 cell line continues to play a pivotal role in advancing our understanding of gene function and developing therapeutic interventions.
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- Antigen expression: SV40 T antigen
- Viruses: SV40 (expresses SV40 T antigen)
- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- 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.
- fluidRenewal: 2 to 3 times per week
- 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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