THLE-3 ATCC ® CRL-11233™ Homo sapiens liver/left lobe These

Permits	These permits may be required for shipping this product: Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Organism	Homo sapiens, human
Tissue	liver/left lobe
Cell Type	epithelialimmortalized with SV40 large T antigen
Product Format	frozen
Morphology	epithelial
Culture Properties	adherent
Biosafety Level	2 Cells contain SV-40 viral DNA sequences
Disease	These immortalized human liver cells constitute an in vitro model for the investigation of etiology and pathogenesis of human hepatocellular carcinoma.
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.
Storage Conditions	liquid nitrogen vapor phase

Karyotype	near diploid RefPfeifer 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
Derivation	The 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.
Genes Expressed	The cells express cytokeratin 18 and albumin in early passage, whereas higher-passage cells in logarithmic-phase growth also express cytokeratin 19.
Tumorigenic	NO
Comments	The 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. 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. 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. THLE-2 and THLE-3 cells metabolize benzo[a]pyrene, N-nitrosodimethylamine, and aflatoxin B1 to their ultimate carcinogenic metabolites that adduct DNA, which indicates functional cytochrome P450 pathways. 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. A culture submitted to the ATCC in January of 1993 was found to be contaminated with mycoplasma. Progeny were cured by a 19-day treatment with Progeny were cured by a 21-day treatment with mycoplasma removal agent (MRA). 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.

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.
Subculturing	Protocol: Remove medium, add fresh 0.05% trypsin - 0.53 mM EDTA, rinse and remove trypsin. Allow the culture to sit at room temperature (or 37C) until the cells detach (about 5 to 15 minutes) . Neutralize the trypsin with 0.1% soybean trypsin inhibitor. Resuspend the cells in fresh medium, aspirate and dispense into coated flasks. Flasks used to propagate these cells must be coated (2 hours at 37C) with 0.01 mg/ml bovine serum albumin, 0.01 mg/ml fibronectin and 0.03 mg/ml bovine collagen type I . Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended Medium Renewal: Every 2 to 3 days
Cryopreservation	Freeze medium: Complete growth medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor phase
Culture Conditions	Temperature: 37°C

STR Profile	Amelogenin: X CSF1PO: 11,12 D13S317: 13 D16S539: 11,12 D5S818: 13 D7S820: 8,10 THO1: 8,9.3 TPOX: 6,9 vWA: 17,18

Name of Depositor	National Cancer Institute
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 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

Name of Depositor

National Cancer Institute

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

Permits

These permits may be required for shipping this product:

Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.

Basic Documentation

Product Sheet

Certificate of Analysis

MSDS

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

THLE-3 (ATCC® CRL-11233™)

Share this product

Your session will expire in

THLE-3 (ATCC^® CRL-11233^™)