UV24 (UV sensitive mutant of CHO) (ATCC® CRL-1866)

Organism: Cricetulus griseus, hamster, Chinese  /  Tissue: ovary  / 

Organism Cricetulus griseus, hamster, Chinese
Tissue ovary
Product Format frozen
Morphology epithelial-like
Culture Properties monolayer and suspension
Biosafety Level 1
Gender female
Applications
The line is defective in nucleotide excision repair, is sensitive to bulky adduct mutagens and belongs to excision repair complementation group 3.
Storage Conditions liquid nitrogen vapor phase
Derivation
This line is a derivative of the CHO-K1 cell line (see ATCC CCL-61).
UV24 is a UV sensitive line derived from AA8 (see ATCC CRL-1859).
Clinical Data
female
Comments
This line is a derivative of the CHO-K1 cell line (see ATCC CCL-61).
UV24 is a UV sensitive line derived from AA8 (see ATCC CRL-1859).
The line is defective in nucleotide excision repair, is sensitive to bulky adduct mutagens and belongs to excision repair complementation group 3.
Complete Growth Medium Alpha minimum essential medium without ribonucleosides and deoxyribonucleosides, 90%; fetal bovine serum, 10%
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. The suspended cells are viable and can be used to start new cultures.

  1. To subculture attached cells, 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.
  2. Observe cells under an inverted microscope until cell layer is dispersed.
    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.
  3. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting. 
  4. Add appropriate aliquots of cell suspension to new culture vessels.
  5. Place culture vessels in incubators at 37°C.

Subcultivation Ratio: 1:4 to 1:12
Medium Renewal: Two times weekly

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.  

Cryopreservation

Complete growth medium described above supplemented with 5% (v/v) DMSO.  Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Population Doubling Time 13 hrs
Name of Depositor LH Thompson
References

Thompson LH, et al. Repair of DNA adducts in asynchronous CHO cells and the role of repair in cell killing and mutation induction in synchronous cells treated with 7-bromomethylbenz[a]anthracene. Somatic Cell Mol. Genet. 10: 183-194, 1984. PubMed: 6584989

Thompson LH, et al. Genetic diversity of UV-sensitive DNA repair mutants of Chinese hamster ovary cells. Proc. Natl. Acad. Sci. USA 78: 3734-3737, 1981. PubMed: 6943579

Hoy CA, et al. Defective DNA cross-link removal in Chinese hamster cell mutants hypersensitive to bifunctional alkylating agents. Cancer Res. 45: 1737-1743, 1985. PubMed: 3919945

Busch D, et al. Summary of complementation groups of UV-sensitive CHO cell mutants isolated by large-scale screening. Mutagenesis 4: 349-354, 1989. PubMed: 2687628

Reardon JT, et al. Isolation and characterization of two human transcription factor IIH (TFIIH)-related complexes: ERCC2/CAKand TFIIH. Proc. Natl. Acad. Sci. USA 93: 6482-6487, 1996. PubMed: 8692841

Thompson LH, et al. Hypersensitivity to mutation and sister-chromatid-exchange induction in CHO cell mutants defective in incising DNA containing UV lesions. Somatic Cell Genet. 8: 759-773, 1982. PubMed: 7163954

. Cellular responses to DNA damage. New York: Liss; 1983.

Bessho T, et al. Initiation of DNA interstrand cross-link repair in humans: the nucleotide excision repair system makes dual incisions 5" to the cross-linked base and removes a 22- to 28-nucleotide-long damage-free strand. Mol. Cell. Biol. 17: 6822-6830, 1997. PubMed: 9372913

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Basic Documentation
References

Thompson LH, et al. Repair of DNA adducts in asynchronous CHO cells and the role of repair in cell killing and mutation induction in synchronous cells treated with 7-bromomethylbenz[a]anthracene. Somatic Cell Mol. Genet. 10: 183-194, 1984. PubMed: 6584989

Thompson LH, et al. Genetic diversity of UV-sensitive DNA repair mutants of Chinese hamster ovary cells. Proc. Natl. Acad. Sci. USA 78: 3734-3737, 1981. PubMed: 6943579

Hoy CA, et al. Defective DNA cross-link removal in Chinese hamster cell mutants hypersensitive to bifunctional alkylating agents. Cancer Res. 45: 1737-1743, 1985. PubMed: 3919945

Busch D, et al. Summary of complementation groups of UV-sensitive CHO cell mutants isolated by large-scale screening. Mutagenesis 4: 349-354, 1989. PubMed: 2687628

Reardon JT, et al. Isolation and characterization of two human transcription factor IIH (TFIIH)-related complexes: ERCC2/CAKand TFIIH. Proc. Natl. Acad. Sci. USA 93: 6482-6487, 1996. PubMed: 8692841

Thompson LH, et al. Hypersensitivity to mutation and sister-chromatid-exchange induction in CHO cell mutants defective in incising DNA containing UV lesions. Somatic Cell Genet. 8: 759-773, 1982. PubMed: 7163954

. Cellular responses to DNA damage. New York: Liss; 1983.

Bessho T, et al. Initiation of DNA interstrand cross-link repair in humans: the nucleotide excision repair system makes dual incisions 5" to the cross-linked base and removes a 22- to 28-nucleotide-long damage-free strand. Mol. Cell. Biol. 17: 6822-6830, 1997. PubMed: 9372913

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.