BT-549 Cells

Name: CLS Cell Lines Service GmbH
Address: Dr.-Eckener-Straße 8, Eppelheim, 69214, DE
Telephone: +496221405780
Price range: €€€

CAD$545.10*

Products are shipped frozen on dry ice in cryotubes. Each cryotube typically contains 3 × 10⁶ cells for adherent lines or 5 × 10⁶ cells for suspension lines (refer to the batch CoA for details).

Prices excl. taxes plus shipping costs

cryovial

Product number: 300132

Safety data sheet

Product sheet

Certificate of Analysis (CoA)

Description	BT-549 cells are a human breast cancer cell line derived from the mammary gland tissue of a 72-year-old Caucasian woman with ductal carcinoma. They are commonly utilized in cancer research to study the biology and treatment of breast cancer, particularly the triple-negative subtype, which lacks estrogen receptor, progesterone receptor, and HER2 expression. BT-549 cells are characterized by their epithelial morphology and are known for their highly invasive properties, making them a valuable model for studying metastasis and tumor invasion. They exhibit several distinctive features including the presence of lipid droplets in the cytoplasm and a robust expression of the mucin-1 protein. These cells also express various oncogenes and tumor suppressor genes that are relevant to breast cancer pathology, such as TP53 and RB1. The BT-549 cell line is estrogen receptor-negative, progesterone receptor-negative, and does not amplify HER2, thus categorizing it under the triple-negative breast cancer (TNBC) subtype. Due to this classification, BT-549 cells are particularly useful for studying the unique mechanisms of progression and treatment response in TNBC, which is known for its aggressive nature and lack of targeted therapies. Furthermore, BT-549 cells are often used in drug resistance studies and for testing new chemotherapeutic agents and targeted therapies, offering insights into potential therapeutic strategies for managing and treating aggressive forms of breast cancer.
Organism	Human
Tissue	Breast, mammary gland
Disease	Invasive ductal carcinoma
Metastatic site	Ductal
Synonyms	BT 549, BT.549, BT549

Age	72 years
Gender	Female
Ethnicity	Caucasian
Morphology	Epithelial-like
Growth properties	Monolayer, adherent

Citation	BT-549 (Cytion catalog number 300132)
Biosafety level	1
NCBI_TaxID	9606
CellosaurusAccession	CVCL_1092

Isoenzymes	G6PD, B, PGM1, 2, PGM3, 1, ES-D, 1, Me-2, 1, AK-1, 1, GLO-1, 1-2, Phenotype Frequency Product: 0.0048
Mutational profile	TP53 mut
Karyotype	Mode = 74, range = 53 to 140, three marker chromosomes

Culture Medium	DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
Supplements	Supplement the medium with 10% FBS
Dissociation Reagent	Accutase
Subculturing	Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
Seeding density	1 x 10⁴ cells/cm² will yield in a confluent layer in about 4 days
Fluid renewal	2 to 3 times per week
Post-Thaw Recovery	After thawing, plate the cells at 5 x 10⁴ cells/cm² and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
Freeze medium	As a cryopreservation medium, we use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
Thawing and Culturing Cells	Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.
Incubation Atmosphere	37°C, 5% CO₂, humidified atmosphere.
Shipping Conditions	Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.
Storage Conditions	For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Sterility	Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

Lot Number	Certificate Type	Date	Catalog Number
300132-310724	Certificate of Analysis	23. May. 2025	300132
300132-1121SF	Certificate of Analysis	23. May. 2025	300132

BT-549 Cells

General information

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Regulatory Data

Biomolecular Data

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Quality Control & Molecular Analysis

Certificate of Analysis (CoA)