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JB6 Cl 30-7b
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NH Colburn
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Biosafety Level:
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1
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frozen
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Medium & Serum:
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See Propagation
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adherent
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Mus musculus (mouse)
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epithelial
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Organ: skin
Strain: BALB/c
Tissue: epidermis
Disease: normal
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In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
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No
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newborn
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ATCC complete growth medium: Minimum essential medium (Eagle) with 2 mM L-glutamine, 95%; fetal bovine serum, 5%
Temperature: 37.0°C
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Protocol: The cells must be carried at low density to avoid spontaneous transformation.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 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. An inoculation density of 2 x 104 viable cells per 25 sq. cm. flask is recommended or a spilt ratio of 1:6 to 1:8.6. Incubate cultures at 37°C.
Subcultivation Ratio: An inoculation density of 2 X 10 exp4 viable cells per 25 sq. cm. flask is recommended
Medium Renewal: 1 to 2 times per week
Rap the flask smartly against a hard surface to get a single cell suspension.
Add fresh medium, aspirate and dispense into new flasks. Culture at 36C in a humidified atmosphere of 5% CO2 - 95% air.
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22309: Colburn NH, et al. A cell culture assay for tumor-promoter-dependent progression toward neoplastic phenotype: detection of tumor promoters and promotion inhibitors. Teratog. Carcinog. Mutagen. 1: 87-96, 1980. PubMed: 6119803
22427: Colburn NH, et al. Correlation of anchorage-independent growth with tumorigenicity of chemically transformed mouse epidermal cells. Cancer Res. 38: 624-634, 1978. PubMed: 626967
22908: Colburn NH, et al. Tumour promoter induces anchorage independence irreversibly. Nature 281: 589-591, 1979. PubMed: 492322
22959: Bernstein LR, Colburn NH. AP1/jun function is differentially induced in promotion-sensitive and resistant JB6 cells. Science 244: 566-569, 1989. PubMed: 2541502
23279: Colburn NH, et al. Dissociation of mitogenesis and late-stage promotion of tumor cell phenotype by phorbol esters: mitogen-resistant variants are sensitive to promotion. Proc. Natl. Acad. Sci. USA 78: 6912-6916, 1981. PubMed: 6947266
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