TK#1 [Embryonic stem cell line] (CRL-11383)

Organism: Mus musculus, mouse  /  Cell Type: embryonic stem cell  / 

Permits and Restrictions

View Permits

Organism Mus musculus, mouse
Cell Type embryonic stem cell
Product Format frozen
Morphology epithelial
Culture Properties adherent
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 embryo
Applications
The cell line was used to produce mutant mice with germ line transmission for the IRF-2 disruption.
Storage Conditions liquid nitrogen vapor phase
Comments

The construct pMIRF2neoB was introduced into D3 embryonic stem (ES) cells by electroporation to disrupt the IRF-2 locus. 

The cell line generated is deficient for the interferon regulatory factor 2 (IRF-2) gene. 

The cell line was used to produce mutant mice with germ line transmission for the IRF-2 disruption. 

Heterozygous mice were inbred to obtain mice homozygous for the disrupted gene. 

The line should be grown on feeder layers of mitomycin C treated primary mouse embryonic fibroblasts or STO cells (see ATCC 56-X.2, MITC-STO cells).
Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose and supplemented with 0.1 mM 2-mercaptoethanol, 85%; fetal bovine serum, 15%
Subculturing

The line should be grown on feeder layers of irradiated (12,000 rads) or mitomycin C treated (0.01 mg/mL for 90 minutes) primary mouse embryonic fibroblasts or STO cells ( ATCC CRL- 1503 ) Irradiated STO cells available as ATCC 56-X.2. The feeder layer should be plated 24 hours in advance

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: 1:4 to 1:8
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 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 TW Mak
U.S. Patent Number
References

Mak TW, Taniguchi T. Mouse lacking the expression of interferon regulatory factor 2 (IRF-2). US Patent 5,675,059 dated Oct 7 1997

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.
  • USDA APHIS VS 16-6 or 16-6A permit must be obtained and a copy of the permit must be sent to ATCC in advance of shipment. The Application Form VS 16-3 (Import controlled material import or transport organisms or vectors) must be submitted to USDA APHIS Veterinary Services to obtain the VS 16-6 or 16-6A permit.
Basic Documentation
References

Mak TW, Taniguchi T. Mouse lacking the expression of interferon regulatory factor 2 (IRF-2). US Patent 5,675,059 dated Oct 7 1997

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