BT-549 (ATCC® HTB-122)

Organism: Homo sapiens, human  /  Cell Type: Epithelial  /  Tissue: mammary gland; breast  /  Disease: ductal carcinoma

Organism Homo sapiens, human
Tissue
mammary gland; breast
Cell Type Epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease ductal carcinoma
Age 72 years
Gender female
Ethnicity Caucasian
Applications
This cell line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
Karyotype modal number = 78; range = 73 to 80.
The cell line is aneuploid human female, with chromosome counts in the hypertriploid range. The X chromosomes are all abnormal. Normal chromosomes N10 and N13 are clearly under-represented, with chromosomes N2, N12, and N17 also tending to be. Chromosome N8 is over-represented with respect to the copy number of other normal chromosomes, and chromosomes N5 and N18 also usually have more copies than most chromosomes. Four marker chromosomes are found: iso(13q), 10q+, del(X)(q22:), del(11)(p11:).
Images
Derivation
The BT-549 line was isolated in 1978 by W.G. Coutinho and E.Y. Lasfargues. Source tissue consisted of a papillary, invasive ductal tumor which had metastasized to 3 of 7 regional lymph nodes.
Clinical Data
72 years
Caucasian
female
Comments
The established population is polymorphic with epithelial like components and multinucleated giant cells. A mucin-like material was secreted into the medium.
Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: 0.023 IU/ml insulin; fetal bovine serum to a final concentration of 10%.


Phenol Red is a structural mimic for estrogen. Therefore the estrogenic activity of phenol red should be considered in any studies that utilize estrogen-responsive cells in culture (RefBerthois Y, et al. Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proc. Natl. Acad. Sci. USA 83: 2496–2500, 1986. PubMed: 3458212). The Phenol Red-free version of RPMI 1640, is ATCC catalog No. 30-2602.

Subculturing
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:6 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
STR Profile
Amelogenin : X
CSF1PO: 10 , 12
D13S317: 11
D16S539 : 8
D5S818: 11
D7S820: 9, 10
TPOX: 8
THO1: 9.3
vWA : 15
Isoenzymes
AK-1, 1
ES-D, 1
G6PD, B
GLO-I, 1-2
Me-2, 1
PGM1, 2
PGM3, 1
Name of Depositor W Coutinho, EY Lasfargues
Year of Origin 1978
References

Katayose Y, et al. Promoting apoptosis: a novel activity associated with the Cyclin-dependent kinase inhibitor p27. Cancer Res. 57: 5441-5445, 1997. PubMed: 9407946

Littlewood-Evans AJ, et al. The osteoclast-associated protease cathepsin K is expressed in human breast carcinoma. Cancer Res. 57: 5386-5390, 1997. PubMed: 9393764

Basic Documentation
Other Documentation
References

Katayose Y, et al. Promoting apoptosis: a novel activity associated with the Cyclin-dependent kinase inhibitor p27. Cancer Res. 57: 5441-5445, 1997. PubMed: 9407946

Littlewood-Evans AJ, et al. The osteoclast-associated protease cathepsin K is expressed in human breast carcinoma. Cancer Res. 57: 5386-5390, 1997. PubMed: 9393764