0166 - MDA-MB-468

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BCRJ Code 0166
Cell Line MDA-MB-468
Species Homo sapiens
Vulgar Name Human
Tissue Mammary Gland/Breast; Derived From Metastatic Site: Pleural Effusion
Morphology Epithelial
Disease Adenocarcinoma
Growth Properties Adherent
Sex Female
Age/Ethinicity 51 Year /
Derivation The MDA-MB-468 cell line was isolated from a pleural effusion of a 51-year-old Black female patient with metastatic adenocarcinoma of the breast.
Applications This cell line is a suitable transfection host.
DNA Profile Amelogenin: X CSF1PO: 12 D13S317: 12 D16S539: 9 D5S818: 12 D7S820: 8 THO1: 7 TPOX: 8,9 vWA: 18
Tumor Formation: Yes, in nude mice inoculated subcutaneously with 10(7) cells. (Tumors developed within 21 days at 100% frequency (5/5).)
Biosafety 1
Addtional Info Although the tissue donor was heterozygous for the G6PD alleles, the cell line consistently showed only the G6PD A phenotype. There is a G -> A mutation in codon 273 of the p53 gene resulting in an Arg -> His substitution. EGF receptor is present at 1 X 10e6 per cell.
Culture Medium Leibovitz's L-15 Medium contains 2 mM L-glutamine, NO sodium bicarbonate and fetal bovine serum to a final concentration of 10%. Note: The L-15 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO2 and air mixture is detrimental to cells when using this medium for cultivation.
Subculturing Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer PBS without calcium and magnesium 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. 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 2 to 3 times per week
Subculturing Subcultivation Ratio 1:2 to 1:4
Culture Conditions Atmosphere: air, 100% 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 1206: Brinkley BR, et al. Variations in cell form and cytoskeleton in human breast carcinoma cells in vitro. Cancer Res. 40: 3118-3129, 1980. PubMed: 7000337 22429: Siciliano MJ, et al. Mutually exclusive genetic signatures of human breast tumor cell line
Depositors MARILIA MEIRA DIAS; UNIVERSIDADE ESTADUAL DE CAMPINAS
Cellosaurus CVCL_0419