BNL CL.2 (ATCC® TIB-73)

Organism: Mus musculus, mouse  /  Tissue: liver  / 

Organism Mus musculus, mouse
Tissue liver
Product Format frozen
Culture Properties adherent
Biosafety Level 1
Age embryo
Strain BALB/c
Storage Conditions liquid nitrogen vapor phase
Tumorigenic No, the cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
Comments

The line was selected to grow in medium containing ornithine and phenylalanine in place of arginine and tyrosine.

Tested and found negative for ectromelia virus (mousepox).

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 1:6
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
Complete growth medium supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions Temperature: 37°C
Atmosphere: 10% CO2
Name of Depositor P Patek, J Collins, M Cohn
References

Patek PQ, et al. Transformed cell lines susceptible or resistant to in vivo surveillance against tumorigenesis. Nature 276: 510-511, 1978. PubMed: 723934

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

Patek PQ, et al. Transformed cell lines susceptible or resistant to in vivo surveillance against tumorigenesis. Nature 276: 510-511, 1978. PubMed: 723934

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.