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C127:LT; Mammary Tumor; Mouse (Mus musculus) Item ID: ATCCRL1804

Permits and Restrictions

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Organism Mus musculus, mouse
Tissue mammary gland
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2  [Cells contain papovavirus]

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.
Gender female
Strain RIII
Applications The C127:LT cell line was derived from the murine mammary tumor line C127 (see also ATCC CRL-1616, C127I) by transfection with polyoma virus DNA. The cells are useful for supporting replication of vectors containing polyoma virus enhancers and origins of DNA replication.
Storage Conditions liquid nitrogen vapor phase
Derivation The C127:LT cell line was derived from the murine mammary tumor line C127 (see also ATCC CRL-1616, C127I) by transfection with polyoma virus DNA.
Comments The C127:LT cell line was derived from the murine mammary tumor line C127 (see also ATCC CRL-1616, C127I) by transfection with polyoma virus DNA. The cells grow to higher saturation densities than untreated C127 cells and constitutively express the polyoma virus large T antigen. The cells are useful for supporting replication of vectors containing polyoma virus enhancers and origins of DNA replication.
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.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM 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: 1:2 to 1:6Medium Renewal: Every 2 to 3 days

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%
Name of Depositor S Nilsson
References

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.

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