RK3E (ATCC® CRL-1895)

Organism: Rattus norvegicus, rat  /  Cell Type: E1A immortalized  /  Tissue: kidney  /  Disease: normal

Organism Rattus norvegicus, rat
Tissue kidney
Cell Type E1A immortalized
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease normal
Age 7 days
Strain Fischer
Applications
transfection host
Storage Conditions liquid nitrogen vapor phase
Derivation
The RK3E cell line was derived by immortalizing rat kidney cells with the E1A oncogene.
Tumorigenic No
Effects
No, The cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
Comments

The cells can be transformed with the RAS and GLI oncogenes.

The cells have an essentially normal karyotype with the exception of one 5q.

Transformation with RAS or GLI renders the cells tumorigenic.

Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum, which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 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. 
  6. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and  subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.  

Name of Depositor K Kinzler, B Vogelstein, J Ruppert
References

Kinzler KW, et al. Identification of an amplified, highly expressed gene in a human glioma. Science 236: 70-73, 1987. PubMed: 3563490

Kinzler KW, Vogelstein B. The GLI gene encodes a nuclear protein which binds specific sequences in the human genome. Mol. Cell. Biol. 10: 634-642, 1990. PubMed: 2105456

Kinzler KW, et al. The GLI gene is a member of the Kruppel family of zinc finger proteins. Nature 332: 371-374, 1988. PubMed: 2832761

Basic Documentation
References

Kinzler KW, et al. Identification of an amplified, highly expressed gene in a human glioma. Science 236: 70-73, 1987. PubMed: 3563490

Kinzler KW, Vogelstein B. The GLI gene encodes a nuclear protein which binds specific sequences in the human genome. Mol. Cell. Biol. 10: 634-642, 1990. PubMed: 2105456

Kinzler KW, et al. The GLI gene is a member of the Kruppel family of zinc finger proteins. Nature 332: 371-374, 1988. PubMed: 2832761