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CT26.CL25 (ATCC® CRL-2639™)

Organism: Mus musculus, mouse  /  Tissue: colon  /  Disease: carcinoma, derived from colon

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Organism Mus musculus, mouse
Tissue colon
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2  [Cells contain SV40 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.
Disease carcinoma, derived from colon
Strain BALB/c
Applications
The cell line can be used as a model for testing immunotherapy protocols and in studies on the host immune response.
Storage Conditions liquid nitrogen vapor temperature
Derivation
CT26 is an N-nitroso-N-methylurethane-(NNMU) induced, undifferentiated colon carcinoma cell line. It was cloned to generate the cell line designated CT26.WT (ATCC CRL-2638).
CT26.WT was stably transduced with the retroviral vector LXSN that contains the lacZ gene encoding the model tumor associated antigen (TAA), beta-galactosidase (beta-gal).
The cells were grown in G418 for seven days, cloned, and evaluated for beta-gal production.
The lethal subclone CT26.CL25 (ATCC CRL-2639) was selected for use in all in vitro and in vivo studies because of its stable high expression of both beta-gal and the class I molecule H-2 Ld.
Clinical Data
The cell line can be used as a model for testing immunotherapy protocols and in studies on the host immune response.
Antigen Expression
H-2d
Genes Expressed
beta galactosidase (beta-gal),H-2d,The lethal subclone CT26.CL25 (ATCC CRL-2639) was selected for use in all in vitro and in vivo studies because of its stable high expression of both beta-gal and the class I molecule H-2 Ld.
The cell line can be used with CT26.CL25 (ATCC CRL-2639) as a model for testing immunotherapy protocols and in studies on the host immune response.
Cellular Products
beta galactosidase (beta-gal)
Tumorigenic Yes
Effects
Yes, in BALB/c mice. Mice inoculated, subcutaneously, developed lethal tumors at 80% frequency with 10 exp3 cells and at 100% with 10 exp4 cells. Pulmonary metastases developed when mice were inoculated, intravenously, with 10 exp4 cells.
Comments
CT26 is an N-nitroso-N-methylurethane-(NNMU) induced, undifferentiated colon carcinoma cell line. It was cloned to generate the cell line designated CT26.WT (ATCC CRL-2638).
CT26.WT was stably transduced with the retroviral vector LXSN that contains the lacZ gene encoding the model tumor associated antigen (TAA), beta-galactosidase (beta-gal).
The vector is driven by the Moloney murine leukemia virus (MoMuLV) long terminal repeat (LTR) promoter and contains a gene controlling resistance to neomycin transcribed from the SV40 promoter.
The cells were grown in G418 for seven days, cloned, and evaluated for beta-gal production.
The lethal subclone CT26.CL25 (ATCC CRL-2639) was selected for use in all in vitro and in vivo studies because of its stable high expression of both beta-gal and the class I molecule H-2 Ld.
The growth rate and lethality of CT26.CL25 and CT26.WT is virtually identical despite the expression by CT26.CL25 of the model TAA, beta-galactosidase, in normal mice.
The cell line can be used as a model for testing immunotherapy protocols and in studies on the host immune response.
Complete Growth Medium RPMI 1640 medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES and 1.0 mM sodium pyruvate and supplemented with 0.1 mM non-essential amino-acids and 0.4 mg/ml G418, 90%; fetal bovine serum, 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.
Note: Subculture at 80% confluence. Cells pile up and do not become completely confluent.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.03% (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: 1:4 to 1:6
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
Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor temperature
Culture Conditions
Temperature: 37.0°C
Name of Depositor N Restifo
References

Wang M, et al. Active immunotherapy of cancer with a nonreplicating recombinant fowlpox virus encoding a model tumor-associated antigen. J. Immunol. 154: 4685-4692, 1995. PubMed: 7722321

Chen PW, et al. Therapeutic antitumor response after immunization with a recombinant adenovirus encoding a model tumor-associated antigen. J. Immunol. 156: 224-231, 1996. PubMed: 8598466

Irvine KR, et al. Cytokine enhancement of DNA immunization leads to effective treatment of established pulmonary metastases. J. Immunol. 156: 238-245, 1996. PubMed: 8598468

Carroll MW, et al. Highly attenuated modified vaccinia virus Ankara (MVA) as an effective recombinant vector: a murine tumor model. Vaccine 15: 387-394, 1997. PubMed: 9141209

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
Product Sheet
Product Sheet
Certificate of Analysis
Certificate of Analysis
SDS
SDS
References

Wang M, et al. Active immunotherapy of cancer with a nonreplicating recombinant fowlpox virus encoding a model tumor-associated antigen. J. Immunol. 154: 4685-4692, 1995. PubMed: 7722321

Chen PW, et al. Therapeutic antitumor response after immunization with a recombinant adenovirus encoding a model tumor-associated antigen. J. Immunol. 156: 224-231, 1996. PubMed: 8598466

Irvine KR, et al. Cytokine enhancement of DNA immunization leads to effective treatment of established pulmonary metastases. J. Immunol. 156: 238-245, 1996. PubMed: 8598468

Carroll MW, et al. Highly attenuated modified vaccinia virus Ankara (MVA) as an effective recombinant vector: a murine tumor model. Vaccine 15: 387-394, 1997. PubMed: 9141209