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293T Cell Line

General Information
OrganismHomo sapiens, human
Cell Line DescriptionThe 293T cell line was created in the Michele Calos laboratory by transfection of a sub-line of adenovirus-immortalized human embryonic kidney cells with a gene encoding the SV40T-antigen and a neomycin resistance gene. The cell line is able to replicate vectors carrying the SV40 origin of replication. The line also has favourable tissue culture, DNA replication, transfection, gene expression, and protein production properties. It gives high titres when used to produce a number of viral vectors such as lentiviruses and oncoretroviruses.
Tissue of OriginKidney (foetal)
Cell TypeSmall slightly rounded cells growing in a dispersed manner
Product FormatFrozen
Growth ModeAdherent
Shipped inDry ice
Storage Temperature−196°C
Culture Conditions and Handling
SubculturingVolumes are given for a 75 cm2 flask. Proportionally increase or reduce the amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard the culture medium.
  2. Briefly rinse the cell layer with Mg2+/Ca2+ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.05% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 1.0-2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5-15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach can be placed at 37°C to facilitate dispersal.
  4. Add 6.0-8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels. And incubate cultures at 37°C.
Subcultivation RatioThe ratio of 1:3 to 1:8 is recommended.
Fluid RenewalEvery 2-3 days
Freeze MediumComplete growth medium 95%; DMSO, 5%
Culture Temperature37°C
AtmosphereAir, 95%; carbon dioxide (CO2)
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