AB.9 (ATCC® CRL-2298)

Organism: Danio rerio, zebrafish  /  Cell Type: fibroblast  /  Tissue: caudal fin  / 

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Organism Danio rerio, zebrafish
Tissue caudal fin
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Biosafety Level 1
Age adult
Strain AB
Applications
This strain of zebrafish is used for genetic mapping. It was used used by Leonard I. Zon at Harvard to generate the zebrafish YAC genomic library and by Peter Goodfellow at Cambridge, United Kingdom to generate the zebrafish radiation hybrid panel.
Storage Conditions liquid nitrogen vapor phase
Derivation
AB.9 is a fibroblast cell line derived from amputated caudal fins of an adult zebrafish, strain AB.
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: heat-inactivated fetal bovine serum to a final concentration of 15%.
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- 0.53 mM 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 28°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 28°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended
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, 95%; DMSO, 5%
Culture Conditions
Temperature: 28°C
Name of Depositor BH Paw, LI Zon
References

Paw BH, Zon LI. Primary fibroblast cell culture. Methods Cell Biol. 59: 39-43, 1999. PubMed: 9891354

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm

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
References

Paw BH, Zon LI. Primary fibroblast cell culture. Methods Cell Biol. 59: 39-43, 1999. PubMed: 9891354

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm