293T (ATCC® CRL-3216)

Organism: Homo sapiens, human  /  Tissue: embryonic kidney  / 

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Organism Homo sapiens, human
Tissue embryonic kidney
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain Adenovirus]

Biosafety  classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Age fetus
Storage Conditions liquid nitrogen vapor phase
Derivation The 293T cell line, originally referred as 293tsA1609neo, is a highly transfectable derivative of human embryonic kidney 293 cells, and contains the SV40 T-antigen.
Comments This cell line is competent to replicate vectors carrying the SV40 region of replication. It gives high titers when used to produce retroviruses. It has been widely used for retroviral production, gene expression and protein production.
Complete Growth Medium The base medium for this cell line is Dulbecco’s Modified Eagle’s Medium (DMEM) (ATCC 30-2002). To make the complete growth medium, add the following components to the base medium: 10% Fetal Bovine Serum (heat inactivated) (ATCC 30-2020), 2mM L-glutamine (ATCC 30-2214), 1% Penicillin/Streptomycin.
Subculturing Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. 
  1. Remove and discard culture medium. 
  2. Briefly rinse the cell layer with Ca++/Mg++ 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 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 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 may be placed at 37°C to facilitate dispersal. 
  4. Add 6.0 to 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. Incubate cultures at 37°C.

Subcultivation ratio: A subcultivation ratio of 1:3 to 1:8 is recommended.
Medium renewal: Every 2 to 3 days

Cryopreservation Freeze medium:  complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions Temperature: 37°C
Atmosphere:air, 95%; carbon dioxide (CO2)
STR Profile

CSF1PO: 11,12
D13S317: 12,14
D16S539: 9,13
D5S818: 8,9
D7S820: 11
TH01: 7, 9.3
TPOX: 11
vWA: 16,19
Amelogenin: X

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation

DuBridge RB, et al. Analysis of mutation in human cells by using an Epstein-Barr virus shuttle system. Mol. Cell Biol. 7: 379-387, 1987. PubMed: 3031469

Pear WS, et al. Production of high-titer helper-free retroviruses by transient transfection. Proc. Natl. Acad. Sci. USA. 90: 8392-8396, 1993. PubMed: 7690960