0046 - B16-F10

.
BCRJ Code 0046
Cell Line B16-F10
Species Mus musculus
Vulgar Name Mouse; C57Bl/6J
Tissue Skin
Morphology Mixture Of Spindle-Shaped And Epithelial-Like Cells
Disease Melanoma
Growth Properties Adherent
Applications This cell line is a suitable transfection host.
Tumor Formation: Yes, in syngeneic mice
Products MELANIN
Biosafety 1
Culture Medium Dulbecco's Modified Eagle's Medium (DMEM) modified to contain 4 mM L-glutamine, 4500 mg/L glucose, 1 mM sodium pyruvate and fetal bovine serum to a final concentration of 10%.
Subculturing Remove medium, and rinse with PBS without calcium and magnesium. Remove the solution and add an additional 1 to 2 mL of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37°C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks. NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010.
Subculturing Medium Renewal Every 2 to 3 days
Subculturing Subcultivation Ratio 1:10
Culture Conditions Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C
Cryopreservation 95% FBS + 5% DMSO (Dimethyl sulfoxide)
Thawing Frozen Cells SAFETY PRECAUTION: It is strongly recommended to always wear protective gloves, clothing, and a full-face mask when handling frozen vials. Some vials may leak when submerged in liquid nitrogen, allowing nitrogen to slowly enter the vial. Upon thawing, the conversion of liquid nitrogen back to its gas phase may cause the vial to explode or eject its cap with significant force, creating flying debris.
  1. Thaw the vial by gently agitating it in a 37°C water bath. To minimize contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as its contents are thawed and decontaminate it by dipping in or spraying with 70% ethanol. From this point, all operations must be performed under strict aseptic conditions.
  3. For cells sensitive to DMSO, it is recommended to remove the cryoprotective agent immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL of complete culture medium and centrifuge at approximately 125 × g for 5 to 7 minutes.
  4. Discard the supernatant and resuspend the cell pellet in the recommended complete medium (see specific batch information for the appropriate dilution ratio).
  5. Incubate the culture under appropriate atmospheric and temperature conditions (see "Culture Conditions" for this cell line).

NOTE: It is important to avoid excessive alkalinity of the medium during cell recovery. To minimize this risk, it is recommended to place the culture vessel containing the growth medium in the incubator for at least 15 minutes before adding the vial contents. This allows the medium to stabilize at its normal pH (7.0 to 7.6).
References Fidler IJ. Biological behavior of malignant melanoma cells correlated to their survival in vivo. Cancer Res. 35: 218-224, 1975. PubMed: 1109790 Fidler IJ, et al. Tumoricidal properties of mouse macrophages activated with mediators from rat lymphocytes stimulated with concanavalin A. Cancer Res. 36: 3608-3615, 1976. PubMed: 953987 Fidler IJ, Bucana C. Mechanism of tumor cell resistance to lysis by syngeneic lymphocytes. Cancer Res. 37: 3945-3956, 1977. PubMed: 908034 Fidler IJ, Kripke ML. Metastasis results from preexisting variant cells within a malignant tumor. Science 197: 893-895, 1977. PubMed: 887927 Briles EB, Kornfeld S. Isolation and metastatic properties of detachment variants of B16 melanoma cells. J. Natl. Cancer Inst. 60: 1217-1222, 1978. PubMed: 418183 Fidler IJ. Selection of successive tumour lines for metastasis. Nat. New Biol. 242: 148-149, 1973. PubMed: 4512654 Li M, et al. Loss of intracisternal A-type retroviral particles in BL6 melanoma cells transfected with MHC class I genes. J.Gen. Virol. 77: 2757-2765, 1996. PubMed: 8922469
Depositors Instituto Nacional do Cancer, Rio de Janeiro.
ATCC CRL-6475
Cellosaurus CVCL_0159