H4TG (ATCC® CRL-1578)

Organism: Rattus norvegicus, rat  /  Tissue: liver  /  Disease: hepatoma

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Organism Rattus norvegicus, rat
Tissue liver
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease hepatoma
Gender male
Strain AxC
Applications
Phenylalanine hydroxylase is expressed consitutively, and production can be increased by treatment with glucorticoids or cAMP derivatives.
The cells are resistant to 6-thioguanine and sensitive to HAT medium.
They are deficient in hypoxanthine-guanine phosphoribosyl transferase (HGPRT-, HPRT-).
Storage Conditions liquid nitrogen vapor phase
Derivation
This line is a derivative of the H4-II-E-C3 rat hepatoma cell line (see ATCC CRL-1600).
Phenylalanine hydroxylase is expressed consitutively, and production can be increased by treatment with glucorticoids or cAMP derivatives.
Clinical Data
male
Comments
This line is a derivative of the H4-II-E-C3 rat hepatoma cell line (see ATCC CRL-1600).
Phenylalanine hydroxylase is expressed consitutively, and production can be increased by treatment with glucorticoids or cAMP derivatives.
The cells are resistant to 6-thioguanine and sensitive to HAT medium.
They are deficient in hypoxanthine-guanine phosphoribosyl transferase (HGPRT-, HPRT-).
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 0.075 mM 6-thioguanine and 10% fetal bovine serum
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 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 TR Chen
References

Haggerty DF, et al. Phenylalanine hydroxylase in cultured hepatoxytes. I. Hormonal control of enzyme levels. J. Biol. Chem. 248: 223-232, 1973. PubMed: 4348207

Peraino C, et al. Hepatomas in tissue culture compared with adapting liver in vivo. Natl. Cancer Inst. Monogr. 13: 229-245, 1964. PubMed: 14143233

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

Haggerty DF, et al. Phenylalanine hydroxylase in cultured hepatoxytes. I. Hormonal control of enzyme levels. J. Biol. Chem. 248: 223-232, 1973. PubMed: 4348207

Peraino C, et al. Hepatomas in tissue culture compared with adapting liver in vivo. Natl. Cancer Inst. Monogr. 13: 229-245, 1964. PubMed: 14143233

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