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HEK 293 STF; Embryonic Kidney; Human (Homo sapiens) Item ID: ATCCRL3249

Permits and Restrictions

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Organism Homo sapiens, human
Tissue embryonic kidney
Cell Type epithelial-like
Product Format frozen
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 2  [Cells contain adenovirus viral DNA sequences]

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
Applications Assaying canonical Wnt signaling pathway. This is a luciferase reporter cell line that responds to canonical Wnt signaling by expressing firefly luciferase.
Storage Conditions liquid nitrogen vapor phase
Images
Comments
  • This cell line was stably cotransfected with (1) a 7 x LEF/TCF sites, a minimal thymidine kinase promoter and (2) a pSV2-neo resistance plasmid exhibiting G418 resistance. 
  • Luciferase activity reflects the level of canonical Wnt signaling.
Complete Growth Medium

The base medium for this cell line is DMEM F12 Medium, (ATCC® 30-2006™), 80%. To make the complete growth medium, add the following components to the base medium:

  • 200 µg/mL G-418
  • Bovine Calf Serum, Iron Fortified (ATCC® 30-2030™), 20%
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).
  3. Add 1.0 to 2.0 mL of 0.25% Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    4. 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 37.0°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommendedMedium Renewal: 2 to 3 times per weekSeeding Density: 3.0 x 104 to 5.0x104 cells/cm2
Cryopreservation Freeze medium: Complete growth medium, 39%; calf serum, 50%; DMSO, 11%Temperature: liquid nitrogen vapor phase
Culture Conditions Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C
Functional Tests canonical Wnt signaling
Name of Depositor J Nathans
Year of Origin 2002
References

Xu Q, et al. Vascular development in the retina and inner ear: control by Norrin and Frizzled-4, a high-affinity ligand-receptor pair. Cell 116:883-895, 2004. PubMed: 15035989

Permits

These permits may be required for shipping this product to Australia:

  • DAFF Import Permit formerly known as AQIS Import Permit must be obtained and a copy of the permit must be sent to ATCC in advance of shipment.
Basic Documentation Product Sheet Certificate of Analysis SDS
Other Documentation Cell Micrograph
References

Xu Q, et al. Vascular development in the retina and inner ear: control by Norrin and Frizzled-4, a high-affinity ligand-receptor pair. Cell 116:883-895, 2004. PubMed: 15035989

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