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Lenti-X293T cells are derived from the human embryonic kidney HEK293T lineage and have been optimized for high-efficiency lentiviral vector production. As in parental HEK293T cells, the SV40 large T antigen is stably expressed, which supports episomal replication of plasmids carrying the SV40 origin of replication and boosts transient expression from these plasmids. The cells grow as an adherent, epithelial-like monolayer under standard serum-supplemented conditions and reach the high densities needed for large-scale virus production.
The line was selected for strong transfection performance with calcium phosphate, lipid-based and polymer-based reagents, giving consistently higher lentiviral titers than conventional HEK293T populations. The cells take up plasmid DNA efficiently, express transgenes strongly and, when co-transfected with suitable packaging and envelope constructs, effectively assemble and release replication-incompetent lentiviral particles. This makes Lenti-X293T cells a common choice for producing third-generation lentiviral vectors for gene delivery, gene editing and stable cell line engineering.
Beyond virus production, the cells remain a useful HEK293-type host for transient gene expression and high-level production of recombinant proteins. Stable growth and reproducible performance suit both small-scale research and scalable production, provided standard biosafety and vector packaging guidelines for lentiviral systems are followed.
- Protein expression: SV40 large T antigen
- Antigen expression: SV40 large T antigen
- Oncogenes: SV40 large T antigen
- Mutational profile: Not fully characterized; contains integrated adenovirus 5 DNA and SV40 large T antigen construct
- Viruses: Contains adenovirus type 5 DNA; expresses SV40 large T antigen
- Virus susceptibility: Highly permissive for lentiviral production
- Tumorigenic: Tumorigenic in immunocompromised mice (for 293T)
- Ploidy status: Aneuploid, hypotriploid (reported for 293T)
- Karyotype: Aneuploid human karyotype with multiple chromosomal abnormalities (typical for 293T)
- GMO status: Genetically modified (adenovirus type 5 DNA transformation; SV40 large T antigen expression)
- Culture medium: 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: 10% FBS
- Incubation atmosphere: 37°C, 5% CO2, humidified atmosphere
- Dissociation reagent: Accutase
- Doubling time: 20–24 hours
- Subculturing: Split before reaching full confluence; allow up to 48 h for full attachment after thawing
- Split ratio: 1:5 to 1:10
- Seeding density: 2 to 4 × 10⁴ cells/cm²
- Fluid renewal: Every 2–3 days
- Thawing: Cells are shipped on dry ice; confirm the vial is still deeply frozen on delivery. Store immediately below −150°C, or thaw quickly in a 37°C water bath (40–60 seconds, until a small ice clump remains). Working in a flow hood, disinfect the vial with 70% ethanol, transfer the cells into 8 ml of room-temperature culture medium in a 15 ml tube, centrifuge at 200 × g for 5 minutes and discard the supernatant containing freezing medium. Resuspend the cell pellet in complete growth medium and plate.
- Freeze medium: Complete growth medium + 10% DMSO
- Storage: Vapor-phase liquid nitrogen (about −150 to −196°C) for long-term storage; −80°C only as a short interim step before transfer to liquid nitrogen
- Shipping: Shipped on dry ice in insulated packaging (approximately −78°C)
- Safety: Wear protective gloves, clothing and a face mask or safety goggles when moving cryovials to or from liquid nitrogen, as vials may burst on removal.
