BT-474 Cells
General information
Description | In 1978, the BT-474 line was isolated by E. Lasfargues and W.G. Coutinho from a solid, invasive ductal carcinoma of the breast. |
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Organism | Human |
Tissue | Breast, mammary gland |
Disease | Invasive ductal carcinoma |
Metastatic site | Ductal |
Synonyms | Bt-474, BT474 |
Characteristics
Age | 60 years |
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Gender | Female |
Ethnicity | Caucasian |
Morphology | Epithelial-like |
Growth properties | The cells grow in compact, slowly growing multi-layered colonies which rarely become confluent. A confluent monolayer is not formed. |
Identifiers / Biosafety / Citation
Citation | BT-474 (Cytion catalog number 300131) |
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Biosafety level | 1 |
Expression / Mutation
Receptors expressed | HER-2/NEU+, ER+, PR+ |
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Isoenzymes | G6PD, B, PGM3, 1, PGM1, 1, ES-D, 1, Me-2, 0, AK-1, 1, GLO-1, 1, Phenotype Frequency Product: 0.0426 |
Tumorigenic | Yes, in nude mice |
Virus susceptibility | mouse mammary tumor virus (RIII-MuMTV) |
Mutational profile | TP53 mut |
Karyotype | mode = 55, range = 50 to 112, bimodal shift 58 - 59 and 100 in later passages with 3 marker chromosomes |
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, 5 microgram/ml Insulin |
Doubling time | 60 to 80 hours |
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 1:3 is recommended |
Seeding density | 2 x 10^4 cells/cm^2 will yield in a mostly confluent layer in about 4 days |
Fluid renewal | 2 to 3 times per week |
Freezing recovery | Almost 100% recovered cells at >90% viability |
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: 10,11
D13S317: 11
D16S539: 9, 11
D5S818: 11, 13
D7S820: 9, 12
TH01: 7
TPOX: 8
vWA: 15, 16
D3S1358: 17
D21S11: 28, 32.2
D18S51: 13, 18
D8S1179: 10, 12
FGA: 22, 25
D1S1656: 13, 15.3
D2S1338: 19
D12S391: 17, 18
D19S433: 14, 17
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HLA alleles |
A*: 01:01:01, 05:02:01
B*: 07:02:01, 20:03:01
C*: 07:02:01, 16:01:01
DRB1*: 04:01:00, 15:01:00
DQA1*: 01:02:01, 03:03:01
DQB1*: 06:02:01
DPB1*: 04:01:01G, 05:01:01G
E: 01:01:01, 01:03:02
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