HEK293T

HEK 293T, a highly transfectable derivative of the parental HEK 293 cell, stands out as a versatile and powerful tool in the field of biotechnology for the production of recombinant proteins and various types of vaccines.

HEK-293T cells were generated by transfecting embryonic kidney 293 cells with a plasmid encoding the SV40 large T antigen. The original HEK293 cell line was developed from the epithelial cells of human embryonic kidney tissue, with its transformation occurring in what was notably the 293rd experiment conducted by the researchers.

In the realm of vaccine development, the 293T embryonic kidney cells are pivotal for viral vector production, including adenovirus vectors. HEK293T cells, under specific culture conditions, are transfected with vectors carrying adenoviral and retroviral elements, including the SV40 origin of replication, leading to the production of virus-like particles (VLPs).

The VLPs, devoid of viral genetic material, are key in forming the basis of subunit and VLP-based vaccines. The recombinant protein production in 293T cells is facilitated by various transfection methods, with an emphasis on the generation of AP fusion proteins and other protein types that form the antigenic component of vaccines.

The 293T cell line's genome engineering capabilities allow for the customization of expression constructs, further boosting the production of viral vectors. This, coupled with the ability to produce proteins in suspension culture or adherent conditions, makes the 293T cell line a full-stack solution for modern vaccine development.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.