產(chǎn)品名稱 |
PTEN-CaP8 |
商品貨號 |
B216271 |
Organism |
Mus musculus, mouse |
Tissue |
prostate epithelium |
Product Format |
frozen |
Morphology |
epithelial-like |
Culture Properties |
adherent |
Biosafety Level |
2 [PTEN-P8 cells were infected with a retrovirus carrying Cre and Puromycin-resistant cassette]
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 |
prostate cancer |
Age |
10 months |
Gender |
male |
Applications |
PTEN deletion and prostate cancer research |
Storage Conditions |
liquid nitrogen vapor phase |
Karyotype |
Chromosome Count: near 6N (115-129)
Numerical Abnormalities: -1, -4, +10, +14, +15, +16, +17, -Y
Structural Abnormalities: Del(5B), Del(7C), Del(8A3), Del(10A1), Del(13B), Del(14C1), Del(15B) |
Derivation |
Mouse prostate epithelium (PTEN-P8) cells were infected with a retrovirus carrying Cre and Puromycin-resistant cassette. |
Receptor Expression |
androgen receptor + |
Tumorigenic |
yes |
Comments |
PTEN mutations are one of the the most frequent genetic alterations found in human prostate cancers. This cell line, PTEN-CaP8, along with its isogenic partner, PTEN-P8 were generated to better understand the underlying molecular mechanisms of PTEN in prostate cancer progression and control, as well as the signaling pathways controlled by PTEN. PTEN-CaP8 is homozygous for Pten deletion. This cell line was generated from tissue that had not been subjected to hormone ablation therapy and are thus ideal for the study of human refractory prostate cancer formation, as many of the most well-studied human prostate cancer cell lines are from late-stage cancer tissue that has undergone such therapy. PTEN-CaP8 cells have lost the second Pten loxp allele and PTEN function as shown by the loss of PTEN protein expression and increased P-AKT levels. Pten null cells are tumorigenic in both male and female severe combined immunodeficiency mice despite no prior exposure to hormone ablation therapy. Complete loss of PTEN causes decreased p27 and E-cadherin expression. |
Complete Growth Medium |
The base medium for this cell line is Dulbecco’s Modified Eagle’s Medium (ATCC® 30-2002™). To make the complete growth medium, add the following components to 500 ml of the base medium:
Fetal bovine serum (FBS; ATCC® 30-2020™) to a final concentration of 10%
25 µg/mL bovine pituitary extract (BPE)
5 µg/mL human recombinant insulin
6 ng/mL human recombinant epidermal growth factor (EGF)
Note: Do not filter complete medium
|
Subculturing |
Subculture when culture reaches ~70-85% confluence.
A subcultivation ratio of 1:20 to 1:50 is recommended.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 8.0 X 103
to 1.0 x 104 viable cells/cm2 is recommended.
- Incubate cultures at 37°C.
Replace medium with fresh medium every 2 to 3 days. |
Cryopreservation |
Freeze Medium: Complete growth medium, 80%; fetal bovine serum, 10% (v/v) and DMSO, 10% (v/v) |
Culture Conditions |
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
Population Doubling Time |
approximately 18 hours |
Name of Depositor |
J Jiao and H Wu |
Year of Origin |
2004 |
References |
Jiao J, et al. Murine cell lines derived from Pten null prostate cancer show the critical role of PTEN in hormone refractory prostate cancer development. Cancer Res. 67: 6083-6091, 2007. PubMed:17616663
Wang S, et al. Prostate-specific deletion of the murine Pten tumor suppressor gene leads to metastatic prostate cancer. Cancer Cell 4(3): 209-221, 2003. PubMed: 14522255
Roy-Burman P, et al. Genetically defined mouse models that mimic natural aspects of human prostate cancer development. Endocr. Relat. Cancer 11: 225–254, 2004. PubMed: 15163300
Freeman D, et al. PTEN tumor suppressor regulates p53 protein levels and activity through phosphatase-dependent and -independent mechanisms. Cancer Cell 3(2): 117-130, 2003. PubMed: 12620407
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