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Designations:
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BT-20
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Depositors:
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EY Lasfargues
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Biosafety Level:
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1
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Shipped:
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frozen
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Medium & Serum:
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See Propagation
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Growth Properties:
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adherent
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Organism:
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Homo sapiens
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Morphology:
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epithelial
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Source:
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Organ: mammary gland; breast
Disease: carcinoma
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Permits/Forms:
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In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
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Isolation:
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Isolation date: 1958
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Applications:
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BT-20 cells are negative for estrogen receptor, but do express an estrogen receptor mRNA that has deletion of exon 5.
This breast tumor line was established by E.Y. Lasfargues and L. Ozzello in 1958 by isolation and cultivation of cells spilling out of the tumor when it was cut in thin slices.
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Tumorigenic:
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Yes
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Reverse Transcript:
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negative
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Antigen expression:
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HLA A1, Bw16 (+/-)
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DNA Profile (STR):
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Amelogenin: X
CSF1PO: 12
D13S317: 11
D16S539: 11,14
D5S818: 12
D7S820: 10
THO1: 7,9.3
TPOX: 11
vWA: 16,17
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Cytogenetic Analysis:
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Normal chromosomes N3, N4, N9, N13, N14, and X may be absent. The markers der(11)t(11;?)(q25;?) (M1); der(1)t(1;3)(p22?;p13?) (M2); and der(2)t(2;?) (q37;?) (M5) were detected by W.A. Nelson-Rees, et al., Int. J. Cancer 16: 74-85, 1975.
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Isoenzymes:
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AK-1, 1-2
ES-D, 1
G6PD, B
GLO-I, 1-2
PGM1, 1
PGM3, 1
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Age:
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74 years
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Gender:
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female
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Ethnicity:
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Caucasian
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Comments:
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The cells express the WNT3 and the WNT7B oncogenes [PubMed: 8168088].
This breast tumor line was established by E.Y. Lasfargues and L. Ozzello in 1958 by isolation and cultivation of cells spilling out of the tumor when it was cut in thin slices. A mycoplasma contaminant was discovered and eliminated early in 1972.
Growth of BT-20 cells is inhibited by tumor necrosis factor alpha (TNF alpha).
BT-20 cells are negative for estrogen receptor, but do express an estrogen receptor mRNA that has deletion of exon 5.
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Propagation:
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ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
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Subculturing:
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Protocol:
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Remove and discard culture medium.
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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.
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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.
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Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
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Add appropriate aliquots of the cell suspension to new culture vessels.
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Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: 2 to 3 times per week
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Preservation:
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Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
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Related Products:
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Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC inkin?id=30-2003">30-2003
recommended serum:ATCC inkin?id=30-2020">30-2020
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References:
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21405: Sugarman BJ, et al. Recombinant human tumor necrosis factor-alpha: effects on proliferation of normal and transformed cells in vitro. Science 230: 943-945, 1985. PubMed: 3933111
22536: Fogh J, et al. Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J. Natl. Cancer Inst. 58: 209-214, 1977. PubMed: 833871
23079: Lan MS, et al. Polypeptide core of a human pancreatic tumor mucin antigen. Cancer Res. 50: 2997-3001, 1990. PubMed: 2334903
23110: Castles CG, et al. expression of a constitutively active estrogen receptor variant in the estrogen receptor-negative BT-20 human breast cancer cell line. Cancer Res. 53: 5934-5939, 1993. PubMed: 8261406
23113: Huguet EL, et al. Differential expression of human Wnt genes 2, 3, 4, and 7B in human breast cell lines and normal and disease states of human breast tissue. Cancer Res. 54: 2615-2621, 1994. PubMed:8168088
23212: Lasfargues EY, Ozzello L. Cultivation of human breast carcinomas. J. Natl. Cancer Inst. 21: 1131-1147, 1958. PubMed: 13611537
23226: Pollack MS, et al. HLA-A, B, C and DR alloantigen expression on forty-six cultured human tumor cell lines. J. Natl. Cancer Inst. 66: 1003-1012, 1981. PubMed: 7017212
32275: 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
32488: Geiger T, et al. Antitumor activity of a PKC-alpha antisense oligonucleotide in combination with standard chemotherapeutic agents against various human tumors transplanted into nude mice. Anticancer Drug Des. 13: 35-45, 1998. PubMed: 9474241
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