AB.9 Cell Line

Cat No.: NTCC5194101

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