RKO (ATCC® CRL-2577)

Organism: Homo sapiens, human  /  Tissue: colon  /  Disease: Carcinoma

Organism Homo sapiens, human
Tissue colon
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease Carcinoma
Applications
It can be used as the control cell line for investigating the effects of p53 and gadd45 on cellular parameters.
Derivation
RKO is a poorly differentiated colon carcinoma cell line developed by Michael Brattain.
Receptor Expression
urokinase receptor (u-PAR)
Oncogene p53 + (wild type) RefSmith ML, et al. Involvement of the p53 tumor suppressor in repair of u.v.-type DNA damage. Oncogene 10: 1053-1059, 1995. PubMed: 7700629
Tumorigenic Yes
Effects
Yes, in nude mice
Yes, in soft agar
Comments

RKO cells contain wild-type p53 but lack endogenous human thyroid receptor nuclear receptor (h-TRbeta1). The level of p53 protein is higher in RKO (ATCC CRL-2577) cells than in RKO-E6 (ATCC CRL-2578) cells.

The RKO cell line is the parental cell line (isogenic) of RKO-E6 (ATCC CRL-2578) and RKO-A545-1 (ATCC CRL-2579).
Complete Growth Medium The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. 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.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53% (w/v) EDTA solution to remove all traces of serum that 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

Subculture Ratio: 1:3 to 1:12
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

Cryopreservation
culture medium 95%; DMSO, 5%
Culture Conditions
Temperature: 37°C

Atmosphere: air, 95%; carbon dioxide (CO2), 5%
STR Profile
Amelogenin: X
CSF1PO: 8, 10
D13S317: 8, 11
D16S539: 12, 13
D5S818: 11, 13
D7S820: 8, 10
THO1: 6, 10
TPOX:11
vWA: 15, 16, 17
Name of Depositor MC Hollander, AJ Fornace
References

Boyd D, et al. Determination of the levels of urokinase and its receptor in human colon carcinoma cell lines. Cancer Res. 48: 3112-3116, 1988. PubMed: 2835152

Smith ML, et al. Involvement of the p53 tumor suppressor in repair of u.v.-type DNA damage. Oncogene 10: 1053-1059, 1995. PubMed: 7700629

Brattain MG, et al. Heterogeneity of human colon carcinoma. Cancer Metastasis Rev. 3: 177-191, 1984. PubMed: 6437669

Bhat MK, et al. Tumor suppressor p53 is a negative regulator in thyroid hormone receptor signaling pathways. J. Biol. Chem. 272: 28989-28993, 1997. PubMed: 9360971

Smith ML, et al. Involvement of the p53 tumor suppressor in repair of u.v.-type DNA damage. Oncogene 10: 1053-1059, 1995. PubMed: 7700629

Basic Documentation
Other Documentation
References

Boyd D, et al. Determination of the levels of urokinase and its receptor in human colon carcinoma cell lines. Cancer Res. 48: 3112-3116, 1988. PubMed: 2835152

Smith ML, et al. Involvement of the p53 tumor suppressor in repair of u.v.-type DNA damage. Oncogene 10: 1053-1059, 1995. PubMed: 7700629

Brattain MG, et al. Heterogeneity of human colon carcinoma. Cancer Metastasis Rev. 3: 177-191, 1984. PubMed: 6437669

Bhat MK, et al. Tumor suppressor p53 is a negative regulator in thyroid hormone receptor signaling pathways. J. Biol. Chem. 272: 28989-28993, 1997. PubMed: 9360971

Smith ML, et al. Involvement of the p53 tumor suppressor in repair of u.v.-type DNA damage. Oncogene 10: 1053-1059, 1995. PubMed: 7700629