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BT-474 Cells

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T8960
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Description

Cat. No. T8960
Name BT-474 Cells
Description Cells were derived from solid, invasive ductal carcinoma from a breast cancer female patient.
Organism Human (H. sapiens)
Tissue Mammary
Donor History Female, Caucasian, 60, Carcinoma (metastatic site, ductal)
Growth Properties Adherent, Epithelial-like
Storage Condition Vapor phase of liquid nitrogen, or below -130°C.
Product Format Frozen
Intended Use This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
BioSafety II
Certificate of Analysis For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com.
Growth Conditions For optimal cell culture, we recommend using PriCoat™ T25 Flasks (G299) or coating your preferred vessels with Applied Cell Extracellular Matrix (G422). DMEM/F-12 (1:1) Medium (TM510) + 5% FBS (*Regular) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂.

*Do not heat-inactivate

Unpacking and Storage Instructions 1. Visually examine the packaging containers for signs of leakage or breakage.

2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130°C, preferably in liquid nitrogen vapor phase storage, until ready for use.

To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability.

Thawing Protocol 1. Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.

2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.

3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.

4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.

5. Incubate the cells at the recommended conditions.

Subculture Protocol Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent.

1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.

2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment.

3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.

4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.

5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired.

6. Incubate the cells at the recommended conditions.

Cryopreservation We recommend using serum-free CryoGuard™  Freezing Media (TM078) or, if serum is preferred, Cryopreservation Medium (TM024).
Seeding Density (cells/cm2) 20,000
Split Ratio 1:2 to 1:3
Population Doubling Time (h) 60-80

ABM

Founded in 2004, Applied Biological Materials (abm) Inc. has been catalyzing scientific discoveries in Life Sciences and Drug Development for over a decade. Based in Vancouver, Canada, our agility in developing and commercializing novel research tools and our complete portfolio of unique services have made us an invaluable partner to laboratories and research institutions around the world. Driven by a team of passionate and talented scientists, abm is dedicated to empowering the scientific community with the latest innovations for life sciences research and drug development.
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