C6/lacZ7 (ATCC® CRL-2303)

Organism: Rattus norvegicus, rat  /  Cell Type: glial cell  /  Disease: glioma

Organism Rattus norvegicus, rat
Cell Type glial cell
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Disease glioma
Strain outbred
Applications
C6/lacZ7 is a subclone of the C6/lacZ cell line (ATCC CRL-2199) that was established in 1989 from the N-nitrosomethylurea-induced glial rat, outbred strain, C6 cell line.
The cells were cultured in G418 for 14 days, cloned, and evaluated for beta-gal production.
The cells constitutively express the lacZ reporter gene product, E. coli derived beta-gal, as revealed on tissue sections by histochemical stain, and single tumor cells can be identified.
Lymphocytes and other responding cells can be identified by double labeling with antibodies on the same slide.
The contrast between stained cells and background facilitates image analysis.
The depositor has confirmed that the C6/lacZ7 subclone grows in rat brain, and that the lacZ marker can be detected by histochemistry for b-gal.
This is one of few models that permit quantitative analysis of microscopic tumor in the brain.
The beta-gal expression is less stable than for 9L/lacZ. Cells should be used soon after thawing or re-cloned.
Methods for the use of C6/lacZ may be found in Cancer Res.
Storage Conditions liquid nitrogen vapor phase
Derivation
C6/lacZ7 is a subclone of the C6/lacZ cell line (ATCC CRL-2199) that was established in 1989 from the N-nitrosomethylurea-induced glial rat, outbred strain, C6 cell line.
The cells constitutively express the lacZ reporter gene product, E. coli derived beta-gal, as revealed on tissue sections by histochemical stain, and single tumor cells can be identified.
Genes Expressed
beta-galactosidase,The subclone was chosen because it has a more stable expression of the lacZ marker.
Cellular Products
beta-galactosidase
Tumorigenic Yes
Effects
Yes, in the brains of CD Fischer 344 rats
Comments
C6/lacZ7 is a subclone of the C6/lacZ cell line (ATCC CRL-2199) that was established in 1989 from the N-nitrosomethylurea-induced glial rat, outbred strain, C6 cell line.
C6 cells were infected with the BAG replication deficient retroviral vector carrying the E. coli lacZ gene encoding beta-gal and the Tn5 neomycin gene, which confers resistance to G418.
The cells were cultured in G418 for 14 days, cloned, and evaluated for beta-gal production.
To obtain the subclone, C6/lacZ was cloned in 1996 at limiting dilution.
The cells constitutively express the lacZ reporter gene product, E. coli derived beta-gal, as revealed on tissue sections by histochemical stain, and single tumor cells can be identified.
Lymphocytes and other responding cells can be identified by double labeling with antibodies on the same slide.
The contrast between stained cells and background facilitates image analysis.
The subclone was chosen because it has a more stable expression of the lacZ marker. The depositor has confirmed that the C6/lacZ7 subclone grows in rat brain, and that the lacZ marker can be detected by histochemistry for b-gal.
This is one of few models that permit quantitative analysis of microscopic tumor in the brain.
The tumor mimics important features of human brain tumor growth and spread.
The beta-gal expression is less stable than for 9L/lacZ. Cells should be used soon after thawing or re-cloned.
The cloned cells have been shown to retain the marker for up to eleven weeks in culture and up to three weeks in the brain.
Methods for the use of C6/lacZ may be found in Cancer Res. 52: 1018-1025, 1992; ibid., 53: 176-182, 1993.
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:
  • O.1 mM Non-Essential Amino Acids (NEAA)
  • fetal bovine serum to a final concentration of 10%

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:
  • O.1 mM Non-Essential Amino Acids (NEAA)
  • fetal bovine serum to a final concentration of 10%

Subculturing
Protocol: Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks.
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:12 is recommended
Medium Renewal: Every 2 to 3 days
Cryopreservation
Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37.0°C
Name of Depositor LA Lampson
References

Lampson LA, et al. Exploiting the lacZ reporter gene for quantitative analysis of disseminated tumor growth within the brain: use of the lacZ gene product as a tumor antigen, for evaluation of antigenic modulation, and to facilitate image analysis of tumor growth in situ. Cancer Res. 53: 176-182, 1993. PubMed: 8416743

Lampson LA, et al. Disseminating tumor cells and their interactions with leukocytes visualized in the brain. Cancer Res. 52: 1018-1025, 1992. PubMed: 1737331

Lampson LA, et al. A new model permits study of the immune response to individual tumor cells within the brain. Neurology 40: 396-397, 1990.

Dutta T, et al. Robust ability of IFN-gamma to upregulate class II MHC antigen expression in tumor bearing rat brains. J. Neuro-Oncol. 64: 31-44, 2003. PubMed: 12952284

Basic Documentation
References

Lampson LA, et al. Exploiting the lacZ reporter gene for quantitative analysis of disseminated tumor growth within the brain: use of the lacZ gene product as a tumor antigen, for evaluation of antigenic modulation, and to facilitate image analysis of tumor growth in situ. Cancer Res. 53: 176-182, 1993. PubMed: 8416743

Lampson LA, et al. Disseminating tumor cells and their interactions with leukocytes visualized in the brain. Cancer Res. 52: 1018-1025, 1992. PubMed: 1737331

Lampson LA, et al. A new model permits study of the immune response to individual tumor cells within the brain. Neurology 40: 396-397, 1990.

Dutta T, et al. Robust ability of IFN-gamma to upregulate class II MHC antigen expression in tumor bearing rat brains. J. Neuro-Oncol. 64: 31-44, 2003. PubMed: 12952284