BT-20 Cells
General information
Description | 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. TNF-alpha (tumor necrosis factor alpha) inhibits the proliferation of BT-20 cells. BT-20 cells are negative for estrogen receptor, but express an estrogen receptor mRNA that has deletion of exon 5. |
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Organism | Human |
Tissue | Breast, mammary gland |
Disease | Invasive ductal carcinoma |
Synonyms | BT 20, BT20 |
Characteristics
Age | 74 years |
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Gender | Female |
Ethnicity | Caucasian |
Morphology | Epithelial-like |
Growth properties | Monolayer, adherent |
Identifiers / Biosafety / Citation
Citation | BT-20 (Cytion catalog number 300130) |
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Biosafety level | 1 |
Expression / Mutation
Antigen expression | HLA A1, Bw16 (+/-) |
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Isoenzymes | PGM3, 1, PGM1, 1, ES-D, 1, AK-1, 1-2, G6PD, B, GLO-1, 1-2, Phenotype Frequency Product: 0.0115 |
Oncogenes | wnt4 +, wnt7h + |
Tumorigenic | Yes, in nude mice. Forms grade II adenocarcinomas |
Reverse transcriptase | negative |
Mutational profile | TP53 mut |
Karyotype | modal number = 50, many markers with large subtelocentrics most characteristic. (P87) Hyperdiploid with abnormalities including fragmented chromosomes, breaks, secondary constrictions, translocations, submetacentric and telocentric markers |
Handling
Culture Medium | DMEM:Ham's F12, w: 3.1 g/L Glucose, w: 1.6 mM L-Glutamine, w: 15 mM HEPES, w: 1.0 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | 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. |
Split ratio | A ratio of 1:2 to 1:4 is recommended |
Seeding density | 1 x 10^4 cells/cm^2 will yield in a confluent layer in about 6 days |
Fluid renewal | 2 to 3 times per week |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
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. |
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STR profile |
Amelogenin: x,x
CSF1PO: 12
D13S317: 11
D16S539: 11, 14
D5S818: 12
D7S820: 10
TH01: 7, 9, 3
TPOX: 11
vWA: 16, 17
D3S1358: 17
D21S11: 28, 29
D18S51: 17
Penta E: 11, 13
Penta D: 10, 11
D8S1179: 12
FGA: 22, 24
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HLA alleles |
A*: 24:02:01, 24:03:01
B*: 15:01:01, 38:01:01
C*: 03:03:01, 12:03:01
DRB1*: 04:04:01, 13:01:01
DQA1*: 01:03:01, 03:01:01
DQB1*: 03:02:01G, 06:01:01G
E: 01:01, 01:03
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