Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease
inflammatory carcinoma of the breast
Age
33 years old
Gender
female
Ethnicity
Caucasian
Applications
The cells have been used for evaluating drug resistance to HER-2 inhibitors, such as trastuzumab and lapatinib. In addition, UACC-732 has been shown to be a drug resistant cell line to cyclin D kinase 4/6 inhibitor and HER-2 inhibitors.
Storage Conditions
liquid nitrogen vapor phase
Karyotype
range=38-76; modal number=71-75
Images
Derivation
This human breast cancer cell line, UACC-732, was derived from a 33 year-old female with infiltrating ductal inflammatory carcinoma of the breast metastatic to the pleural fluid. Pathology confirmed that the specimen was a metastatic adenocarcinoma consistent with inflammatory carcinoma of the breast.
The base medium for this cell line is ATCC-formulated Leibovitz's L-15 Medium, Catalog No. 30-2008. To make the complete growth medium, add the following components to the base medium:
10 ng/ml human recombinant epidermal growth factor (EGF) (final conc.)
3 ng/ml (0.01 µM) estradiol (final conc.)
0.8 ng/ml (1 pM) Na-L-thyroxine (final conc.)
23 µg/mL (0.2mM) proline (final conc.)
extra 2 mM glutamine
Note: Do not filter complete medium. L-15 Medium is formulated 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
These cells are cultured on collagen I coated vessels. Add 5 µg per cm2 collagen I (BD Biosciences, Cat. No.354236 or equivalent) to culture vessels and incubate at room temperature for 1 hour. Remove collagen solution and rinse vessels 3 times with a balanced salt solution. Vessels may be used immediately or air dried and stored at 2-8°C for up to one week under sterile conditions. Alternatively, commercially available pre-coated Collagen I vessels, such as BD BioCoat Cellware (BD Biosciences, Cat. No. 356524 for 75 cm2 flask) or equivalent, may be used.
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of solutions for culture vessels of other sizes.
Remove and discard culture medium.
Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (DPBS) or 0.25% (w/v) Trypsin - 0.53 mM 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.
Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new collagen I coated culture vessels. An inoculum of 3 X 104 to 4 X 104 viable cells/cm2 is recommended.
Incubate cultures at 37.0°C.
Subculture when cultures reach a cell concentration between 7 X 104 to 1 X 105
Subcultivation ratio: A subcultivation ratio of 1:2 is recommended. Medium renewal: every 2 to 3 days Note: This cell line grows slowly.
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