| 產(chǎn)品名稱 |
TRAMP-C1 |
| 商品貨號(hào) |
B162462 |
| Organism |
Mus musculus, transgenic, mouse, transgenic |
| Tissue |
prostate |
| Cell Type |
epithelial |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain SV40 DNA viral sequences]
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 |
male |
| Strain |
C57BL/6-TgN(TRAMP)8247Ng |
| Applications |
The cell lines can be used in studies to elucidate molecular mechanisms associated with the initiation, progression and metastasis of prostate cancer. They are also a useful tool for gene/drug discovery. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
The TRAMP-C1 (ATCC- CRL-2730), TRAMP- C2 (ATCC-CRL-2731) and TRAMP-C3 (ATCC CRL-2732) cell lines were derived in 1996 from a heterogeneous 32 week primary tumor in the prostate of a PB-Tag C57BL/6 (TRAMP) mouse. |
| Clinical Data |
male |
| Receptor Expression |
androgen receptor, expressed |
| Genes Expressed |
E-cadherin, cytokeratin |
| Cellular Products |
E-cadherin cytokeratin |
| Tumorigenic |
Yes |
| Effects |
Yes, C57BL/6 hosts No, soft agar |
| Comments |
TRAMP is a transgenic line of C57BL/6 mice harboring a construct comprised of the minimal -426/+28 rat probasin promoter (426 base pairs of the rat probasin (PB) gene promoter and 28 base pairs of 5'-untranslated region) to target expression of the SV40 large T antigen to prostatic epithelium. Neither the cells grown in culture, nor the tumors arising from the cells in vivo, express SV40 T antigen (Tag). TRAMP-C1 and TRAMP-C2 are tumorigenic when grafted into syngeneic C57BL/6 hosts. However, TRAMP-C3 grows readily in vitro, but does not form tumors. These cell lines represent various stages of cellular transformation and progression to androgen-independent metastatic disease that can be manipulated in vitro. |
| Complete Growth Medium |
Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplemented with 0.005 mg/ml bovine insulin and 10 nM dehydroisoandrosterone, 90%; fetal bovine serum, 5%; Nu-Serum IV, 5%.
|
| Subculturing |
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- 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 that 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 pipetting gently.
- To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to10 minutes.
- Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
- Place culture vessels in incubators at 37°C.
Subcultivation Ratio: 1:6 to 1:10
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells: a Manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss N.Y., 2005. |
| Cryopreservation |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
| Population Doubling Time |
25 hrs |
| Name of Depositor |
N Greenberg |
| Deposited As |
mouse, transgenic |
| Year of Origin |
1996 |
| References |
Hurwitz AA, et al. Manipulation of T cell costimulatory and inhibitory signals for immunotherapy of prostate cancer. Proc. Natl. Acad. Sci. USA 94: 8099-8103, 1997. PubMed: 9223321
Foster BA, et al. Characterization of prostatic epithelial cell lines derived from transgenic adenocarcinoma of the mouse prostate (TRAMP) model. Cancer Res. 57: 3325-3330, 1997. PubMed: 9269988
Greenberg NM, et al. Prostate cancer in a transgenic mouse. Proc. Natl. Acad. Sci. USA 92: 3439-3443, 1995. PubMed: 7724580
Greenberg NM, et al. The rat probasin gene promoter directs hormonally and developmentally regulated expression of a heterologous gene specifically to the prostate in transgenic mice. Mol. Endocrinol. 8: 230-239, 1994. PubMed: 8170479
Gingrich JR, et al. Metastatic prostate cancer in a transgenic mouse. Cancer Res. 56: 4096-4102, 1996. PubMed: 8797572
Greenberg NM. Transgenic models for prostate cancer research. Urol. Oncol. 2: 119-122, 1996.
Gingrich JR, et al. Androgen-independent prostate cancer progression in the TRAMP model. Cancer Res. 57: 4687-4691, 1997. PubMed: 9354422
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