PY8119 cell line细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

PY8119 cell line细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心
Organism Mus musculus, mouse
Tissue mammary gland
Cell Type mesenchymal-like
Product Format frozen 1.0 mL
Morphology fibroblast-like
Culture Properties adherent
Biosafety Level 1
Derivation Py8119 cells were obtained from spontaneously arising tumors in MMTV-PyMY (mouse mammary tumor virus promoter driven Polyoma middle T-antigen) transgenic C57BL/6 female mice. Tumors were minced and incubated in collagenase, passed through 70 µM nylon mesh and cultured. The cells were subsequently cloned by serial trypsinization and limiting dilution.
Antigen Expression N-cadherin+, vimentin+, cytokeratin 14+, Slug (SNAI2)+ (PubMed: 24368187)
Receptor Expression estrogen receptor, not expressed
progesterone receptor, not expressed
HER2 (human epidermal growth factor receptor 2), not expressed (PubMed: 22531600)
Tumorigenic yes
Effects tail vein injection of Py8119 cells results in tumors in multiple sites including lung, liver and bone
Comments The P8119 cell line represents a mesenchymal tumor cell derived from a mammary adenocarcinoma that spontaneously arose in a MMTV-PyMY (mouse mammary tumor virus promoter driven Polyoma middle T-antigen) transgenic C57BL/6 female mouse. This cell line carries the Polyoma virus middle T oncogene, however its expression is down-regulated. (L Ellies, personal communication)
Py8119 cells are, to a degree, spindle-shaped and do not form discrete colonies in culture. (PubMed: 24368187)
This cell line is very robust and forms aggressive mesenchymal tumors in vivo. The tumors are not metastasized when injected orthotopically, however tail vein injection results in tumors in multiple sites including lung, liver and bone. (L Ellies, personal communication)
Py8119 express mesenchymal markers N-cadherin, vimentin, Slug (SNAI2) and cytokeratin 14 and is negative for expression of estrogen receptor, progesterone receptor and HER2 (human epidermal growth factor receptor 2). (PubMed: 24368187)
The Py8119 cell line exhibits a more undifferentiated phenotype and grows more aggressively in cell culture in comparison to the more differentiated and epithelial-like CRL-3279, Py230 cells, which were also derived from a MMTV-PyMY transgenic C57BL/6 female mouse (PubMed: 22531600)
Py8119 cells, along with Py230 cells, are a pair of murine mammary cell lines with distinct mesenchymal (Py8119) or epithelial-like (Py230) features derived from MMTV-PyMT transgene-induced mammary tumors in C57BL/6 mice that can be useful to study mammary tumorigenesis. (PubMed: 24368187)
Complete Growth Medium The base medium for this cell line is F-12K Medium (BIOVECTOR 30-2004). To make the complete growth medium, add the following component to the 500 mL of the base medium:
• Fetal bovine serum (FBS; BIOVECTOR 30-2020) for a final concentration of 5%
Subculturing Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
1. Remove and discard culture medium. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) (BIOVECTOR 30-2200) or 0.25% (w/v) Trypsin - 0.53 mM EDTA (BIOVECTOR 30-2101) solution to remove all traces of serum which contains trypsin inhibitor.
2. 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.
1. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
2. Transfer cell suspension to a centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes.
3. Discard supernatant. Resuspend the cell pellet in fresh growth medium.
4. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 1 X 104 to 5 X 104 viable cells/cm2 is recommended.
5. Incubate cultures at 37°C. Subculture when cell density reaches between 2 X 105 and 3 X 105 cells/cm2.

Subcultivation Ratio: 1:5 to 1:10 is recommended.
Medium Renewal: 2 to 3 times a week
Cryopreservation Freeze Medium: Complete culture medium + 25% FBS (BIOVECTOR30-2020) + 10% DMSO (BIOVECTOR 4-X)
Storage Temperature: liquid nitrogen vapor phase
Culture Conditions Temperature: 37°C; Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Complete Growth Medium The base medium for this cell line is F-12K Medium (BIOVECTOR 30-2004). To make the complete growth medium, add the following component to the 500 mL of the base medium:
• Fetal bovine serum (FBS; BIOVECTOR 30-2020) for a final concentration of 5%
Subculturing Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
1. Remove and discard culture medium. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) (BIOVECTOR 30-2200) or 0.25% (w/v) Trypsin - 0.53 mM EDTA (BIOVECTOR 30-2101) solution to remove all traces of serum which contains trypsin inhibitor.
2. 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.
1. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
2. Transfer cell suspension to a centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes.
3. Discard supernatant. Resuspend the cell pellet in fresh growth medium.
4. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 1 X 104 to 5 X 104 viable cells/cm2 is recommended.
5. Incubate cultures at 37°C. Subculture when cell density reaches between 2 X 105 and 3 X 105 cells/cm2.

Subcultivation Ratio: 1:5 to 1:10 is recommended.
Medium Renewal: 2 to 3 times a week
Cryopreservation Freeze Medium: Complete culture medium + 25% FBS (BIOVECTOR30-2020) + 10% DMSO (BIOVECTOR 4-X)
Storage Temperature: liquid nitrogen vapor phase
Culture Conditions Temperature: 37°C; Atmosphere: air, 95%; carbon dioxide (CO2), 5%

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