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微信小章| 产品名称： | Py230 |
|---|---|
| 商品货号： | TS205466 |
| Organism： | Mus musculus, mouse |
| Tissue： | mammary gland |
| Cell Type： | epithelial-like |
| Product Format： | frozen 1.0 mL |
| Morphology： | epithelial-like |
| Culture Properties： | adherent |
| Biosafety Level： | 1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease： | adenocarcinoma |
| Age： | adult |
| Gender： | female |
| Applications： | This cell line is a model for investigating the multistep progression of malignant mammary tumorigenesis and metastasis and can also be used as a precllinical mouse model of triple negative breast cancer. |
| Storage Conditions： | liquid nitrogen vapor phase |
| Images： |  |
| Derivation： | Py230 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： | E-cadherin+ (PubMed: 24368187), cytokeratin 8+, cytokeratin 14+ |
| Receptor Expression： | estrogen receptor, not expressed, progesterone receptor, not expressed HER2 (human epidermal growth factor receptor 2), low expression (PubMed: 22531600) |
| Tumorigenic： | yes |
| Effects： | forms luminal tumors and lung metastasis |
| Comments： | The Py230 cell line is an epithelial-like murine mammary luminal tumor cell line that has properties similar to those of normal mouse mammary stem cells. It was 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. (L Ellies, personal communication) Py230 cells are cuboidal-shaped and grow in well-differentiated colonies at confluence in culture. (PubMed:xa0 24368187) This cell line forms luminal tumors and lung metastasis in vivo. (Deposit Form) Py230 cells differentiate into single positive luminal, myoepithelial, and alveolar cells upon stimulation with lactogenic hormones. (L Ellies, personal communication) Py230 cells express luminal-epithelial markers E-cadherin, cytokeratin 8 and cytokeratin14. Py230 cells are negative for expression of estrogen receptor, progesterone receptor and express low levels of HER2. (PubMed:xa0 24368187)xa0 The Py230 cell line has a more differentiated phenotype and grows less aggressively in cell culture in comparison to the undifferenitiated and mesenchymal CRL-3280, Py8119 cells, which were also derived from a MMTV-PyMY transgenic C57BL/6 female mouse. (PubMed:xa0 22531600) Py230 cells, along with Py8119 cells, are a pair of murine mammary cell lines with distinct epithelial-like (Py230) or mesenchymal (Py8119) features derived from MMTV-PyMT transgene-induced mammary tumors in C57BL/6 mice that can be useful to study mammary tumorigenesis. (PubMed:xa0 24368187)xa0 |
| Complete Growth Medium： | The base medium for this cell line is F-12K Medium (ATCC 30-2004). To make the complete growth medium, add the following component to the 500 mL of the base medium:
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| Subculturing： | Note: these cells will not maintain their differentiation properties without MITO+ Serum Extender. Note: these cells grow slowly 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 Dulbeccos phosphate-buffered saline (D-PBS) (ATCC 30-2200) or 0.25% Trypsin – 2.21mM EDTA in HBSS (Corning cat no. 25-053-Cl) 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). xa0xa0xa0xa0Note: xa0These cells will form small clumps upon trypsinization.They do not form a single cell suspension. 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. 3. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. An inoculum of 3 X 104 to 5 X 104 viable cells/cm2 is recommended. 4. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Subculture when cell density reaches between 2 X 105 and 3 X 105 cells/cm2. xa0 xa0 Subcultivation Ratio: 1:3 to 1:6 is recommended. Medium renewal: every other day xa0 Note: These cells will differentiate if maintained at low density. Note: It is recommended to make clones from early passages. Subclone by plating into ultra-low adhesion plates at low density for 2 days. Lightly trypsinize cells and pick single cells, place into wells of 96-well plate with 150µL complete medium. Leave in incubator for 2 weeks. Check for colony growth. Keep only colonies that grow and form domes. |
| Cryopreservation： | Freeze Medium: Complete culture medium + 40% additional FBS (ATCC 30-2020) + 10% DMSO (ATCC 4-X) Storage Temperature: liquid nitrogen vapor phase |
| Culture Conditions： | Temperature: 37°C Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| Volume： | 1.0 mL |
| Name of Depositor： | L Ellies, University of California, San Diego |
| Year of Origin： | June 2004 |
| References： | Guy CT, et al. Induction of mammary tumors by expression of polyomavirus middle T oncogene: a transgenic mouse model for metastatic disease. Mol. Cell Biol. 12: 954-961, 1992. PubMed: 1312220 Maglione JE, et al. Transgenic Polyoma middle-T mice model premalignant mammary disease. Cancer Res. 61: 8298-8305, 2001. PubMed: 11719463 Gibby K, et al. Early vascular deficits are correlated with delayed mammary tumorigenesis in the MMTV-PyMT transgenic mouse following genetic ablation of the NG2 proteoglycan. Breast Cancer Res. 14: R67, 2012. PubMed: 22531600 Biswas T, et al. Attenuation of TGF-β signaling supports tumor progression of a mesenchymal-like mammary tumor cell line in a syngeneic murine model. Cancer Lett 346: 129-138, 2014. PubMed: 24368187 Bao L, et al. Multipotent luminal mammary cancer stem cells model tumor heterogeneity. Breast Cancer Res. 17(1): 137, 2015. PubMed: 26467658 |