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AB.9
AB.9
規格:
價格:詢價
編號:B163901
品牌:Mingzhoubio

標準菌株
定量菌液
DNA
RNA

規格:
凍干粉
斜面
甘油
平板


產品名稱
AB.9
商品貨號
B163901
Organism
Danio rerio, zebrafish
Tissue
caudal fin
培養基
Cell Type
fibroblast
Product Format
frozen
Morphology
fibroblast
Biosafety Level
1

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.

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-0.5% PVP solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of 0.25% (w/v) Trypsin-0.53 mM EDTA-0.5% PVP 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.

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