品系:C57L/J
ATCC Number:CRL-2711?
相關(guān)疾?。焊伟?br />
運(yùn)輸方式:凍存運(yùn)輸
細(xì)胞形態(tài):上皮樣
細(xì)胞類型:上皮細(xì)胞
生長狀態(tài):貼壁生長
是否是腫瘤細(xì)胞:0
物種來源:小鼠
數(shù)量:大量
器官來源:肝
規(guī)格:0.1ml Designations: c37 (B7IFi1)
Depositors: ?O Hankinson
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism:
Mus musculus deposited as mouse
Morphology:epithelial
Source:
Organ: liver
Strain: C57L/J
Disease: hepatoma
Cell Type: epithelial
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Comments:The c37 (B7IFi1) cell line was derived from Hepa-1c1c7 (ATCC CRL-2026 ). Hepa-1c1c7 has high aryl hydrocarbon hydroxylase (AHH) activity. ICR191G (a frame shift mutagen) was used to treat Hepa-1c1c7 cells. Mutated colonies were selected for benzo[a]pyrene resistance. The c37 (B7IFi1) cell line is one of these, and lacks cytochrome P4501A1 dependent aryl hydrocarbon hydroylase (AHH) activity due to two point mutations in the CYP1A1 protein (Leu-118 to Arg and Arg-245 to Pro). The line may be used to study xenobiotic (and carcinogenic) metabolism in the absence of cytochrome P4501A1 activity, which is known to metabolize benzo[a]pyrene cytotoxic and carcinogenic intermediates. It is also a tool to study the putative natural ligand for the induction of this enzyme.
Propagation:
ATCC complete growth medium: Alpha minimum essential medium without ribonucleosides and deoxyribonucleosides with 2 mM L-glutamine, 90%; heat-inactivated fetal bovine serum, 10%
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing:
Protocol:
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Remove and discard culture medium.
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Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
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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.
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Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
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Add appropriate aliquots of the cell suspension to new culture vessels.
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Incubate cultures at 37?C.
Subcultivation Ratio: A subcultivation ratio of 1:5 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days
Preservation:
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products:parental cell line:ATCC CRL-2026
References: 22593: Hankinson O. Single-step selection of clones of a mouse hepatoma line deficient in aryl hydrocarbon hydroxylase. Proc. Natl. Acad. Sci. USA 76: 373-376, 1979. PubMed: 106390
61151: Kimura S, et al. Analysis of two benzo[a]pyrene-resistant mutants of the mouse hepatoma Hepa-1 P(1)450 gene via cDNA expression in yeast. EMBO J. 6: 1929-1933, 1987. PubMed: 3308449