| Applications |
This cell line along with the wild-type Hepa-1clc7 and the class I variant tao BpRcl (see ATCC CRL-2218) can be used to study dioxin (TCDD) induction of the CYPIA1 gene, which precedes increased levels of p450IA1 enzyme. BpRc1 is a Class II variant of the Hepa-1clc7 (see ATCC CRL-2026) hepatoma cell line which was derived from the Hepa-1 cell line established by Bernhard, et al. |
| Derivation |
BpRc1 is a Class II variant of the Hepa-1clc7 (see ATCC CRL-2026) hepatoma cell line which was derived from the Hepa-1 cell line established by Bernhard, et al. This cell line along with the wild-type Hepa-1clc7 and the class I variant tao BpRcl (see ATCC CRL-2218) can be used to study dioxin (TCDD) induction of the CYPIA1 gene, which precedes increased levels of p450IA1 enzyme. The p450IAI enzyme catalyzes hydroxylase activity which results in the oxygenation of aromatic substrates such as the environmental carcinogen benzo[a]pyrene. Induction of transcription requires the aryl hydrocarbon hydroxylase (Ah) receptor to bind TCDD and transport of the TCDD receptor complex into the nucleus where interaction with a dioxin responsive enhancer (DRE) upstream of the CYPIA1 gene facilitates increased transcription. |
| Comments |
BpRc1 is a Class II variant of the Hepa-1clc7 (see ATCC CRL-2026) hepatoma cell line which was derived from the Hepa-1 cell line established by Bernhard, et al. This cell line along with the wild-type Hepa-1clc7 and the class I variant tao BpRcl (see ATCC CRL-2218) can be used to study dioxin (TCDD) induction of the CYPIA1 gene, which precedes increased levels of p450IA1 enzyme. The p450IAI enzyme catalyzes hydroxylase activity which results in the oxygenation of aromatic substrates such as the environmental carcinogen benzo[a]pyrene. Induction of transcription requires the aryl hydrocarbon hydroxylase (Ah) receptor to bind TCDD and transport of the TCDD receptor complex into the nucleus where interaction with a dioxin responsive enhancer (DRE) upstream of the CYPIA1 gene facilitates increased transcription. |
| 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.
- Remove and discard culture medium.
- 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.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: 1:4 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. |
| References |
Bernhard HP, et al. Expression of liver phenotypes in cultured mouse hepatoma cells: synthesis and secretion of serum albumin. Dev. Biol. 35: 83-96, 1973. PubMed: 4362668
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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