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微信小章| 产品名称： | THLE-2 |
|---|---|
| 商品货号： | TS210752 |
| Organism： | Homo sapiens, human |
| Tissue： | liver/left lobe |
| Cell Type： | epithelial cells transformed with SV40 large T antigen |
| Product Format： | frozen |
| Morphology： | epithelial |
| Culture Properties： | adherent |
| Biosafety Level： | 2 Cells contain SV40 viral DNA 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. |
| Age： | adult |
| Applications： | These immortalized human liver cells constitute an in vitro model for pharmacotoxicological studies and for the investigation of etiology and pathogenesis of human hepatocellular carcinoma. xad
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| Storage Conditions： | liquid nitrogen vapor phase |
| Disclosure： | This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC. |
| Karyotype： | near diploid |
| Images： |  |
| Derivation： | TThe THLE-2 (ATCC CRL-10149 and the THLE-3 (ATCC CRL-11233) cell lines were derived from primary normal liver cells by infection with SV40 large T antigen. RF84749 The virus was generated by introducing a retroviral vector containing the of Bgl I-Hpa I fragment of SV40 T antigen into the amphotropic packaging cell line PA317. |
| Tumorigenic： | No |
| Effects： | No, nude mice |
| Comments： | RF84750 THLE-2 and THLE-3 cells express phenotypic characteristics of normal adult liver epithelial cells. They are nontumorigenic when injected into athymic nude mice, have near-diploid karyotypes, and do not express alpha-fetoprotein. RF84750 THLE-2 and THLE-3 cells metabolize benzoapyrene, N-nitrosodimethylamine, and aflatoxin B1 to their ultimate carcinogenic metabolites that adduct DNA, which indicates functional cytochrome P450 pathways. RF84750 Other enzymes involved in metabolism of chemical carcinogens, such as epoxide hydrolase, NADPH cytochrome P450 reductase, superoxide dismutase, catalase, glutathione S-transferases, and glutathione peroxidase are also retained by THLE cells. RF84750 xa0A culture submitted to the ATCC in May of 1989 was found to be contaminated with mycoplasma. Progeny were cured by a 21-day treatment with BM Cycline. The cells were assayed for mycoplasma, by the Hoechst stain, PCR and the standard culture test, after a six-week period following treatment. All tests were negative. The cured cell line is available as TS210752. The original patent deposit is available as ATCC CRL-10149 |
| Complete Growth Medium： | BEGM from Lonza/Clonetics Corporation, Walkersville, MD 21793 (BEGM Bullet Kit; CC3170). The kit includes 500 mL basal medium and separate frozen additives from which we discard the gentamycin/ Amphotericin (GA) and Epinephrine and to which we add extra 5 ng/mL EGF, 70 ng/mL Phosphoethanolamine and 10% fetal bovine serum.
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| 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.
Note: The flasks used should be precoated with a mixture of 0.01 mg/mL fibronectin, 0.03 mg/mL bovine collagen type I and 0.01 mg/mL bovine serum albumin dissolved in BEBM medium.xa0
Subculture Ratio: 1:3 to 1:6 Medium Renewal: Two to three times a week.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994. |
| 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 |
| STR Profile： | Amelogenin: X, Y CSF1PO: 11, 13 D13S317: 8, 12 D16S539: 11, 13 D5S818: 11, 13 D7S820: 10, 12 TH01: 7, 9.3 TPOX: 8, 11 vWA: 16, 17 |
| Name of Depositor： | National Cancer Institute |
| Deposited As： | human |
| U.S. Patent Number： | |
| References： | Harris CC, et al. Human liver epithelial cells. US Patent 5,759,765 dated Jun 2 1998 Pfeifer AM, et al. Simian virus 40 large tumor antigen-immortalized normal human liver epithelial cells express hepatocyte characteristics and metabolize chemical carcinogens. Proc. Natl. Acad. Sci. USA 90: 5123-5127, 1993. PubMed: 7685115 |