產(chǎn)品名稱 |
Het-1A |
商品貨號(hào) |
B164599 |
Organism |
Homo sapiens, human |
Tissue |
esophagus |
Cell Type |
epithelial; SV40 large T antigen transfected |
Product Format |
frozen |
Morphology |
epithelial |
Culture Properties |
adherent |
Biosafety Level |
2 [cells contain SV40 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 |
normal |
Age |
25 years |
Gender |
male |
Ethnicity |
Black |
Applications |
The cell line may be useful for investigating the action of putative esophageal carcinogens. |
Storage Conditions |
liquid nitrogen vapor phase |
Karyotype |
hypodiploid (34-40 chromosomes)
RefStoner GD, et al. Establishment and characterization of SV40 T-antigen immortalized human esophageal epithelial cells. Cancer Res. 51: 365-371, 1991. PubMed: 1703038 |
Derivation |
The HET-1A cell line was derived in 1986 from human esophageal autopsy tissue by transfection with plasmid pRSV-T consisting of the RSV-LTR promoter and the sequence encoding the simian virus 40 large T-antigen. |
Clinical Data |
25 years
Black
male |
Antigen Expression |
SV40 T antigen |
Genes Expressed |
cytokeratin Ref Stoner GD, et al. Establishment and characterization of SV40 T-antigen immortalized human esophageal epithelial cells. Cancer Res. 51: 365-371, 1991. PubMed: 1703038 |
Cellular Products |
cytokeratin Ref Stoner GD, et al. Establishment and characterization of SV40 T-antigen immortalized human esophageal epithelial cells. Cancer Res. 51: 365-371, 1991. PubMed: 1703038 |
Tumorigenic |
No |
Effects |
No, nontumorigenic in athymic, nude mice for more than 12 months. Ref Stoner GD, et al. Establishment and characterization of SV40 T-antigen immortalized human esophageal epithelial cells. Cancer Res. 51: 365-371, 1991. PubMed: 1703038 |
Comments |
The line has undergone more than 250 population doublings.
Growth factor studies have shown that HET-1A is stimulated by calcium and inhibited by fetal bovine serum, transforming growth factor-beta 1, and transforming growth factor-beta 2.
Fumonisin B1 produces growth inhibition and increased apoptosis in HET-1A cells.
The synthetic retinoid CD437 (6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid (AHPN/CD437)0 induces apoptosis of esophageal squamous HET-1A cells through the caspase-3-dependent pathway.
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Complete Growth Medium |
The base medium for this cell line (BEBM) along with all the additives can be obtained from Lonza/Clonetics Corporation as a kit: BEGM, Kit Catalog No. CC-3170. ATCC does not use the GA-1000 (gentamycin-amphotericin B mix) provided with the BEGM kit. Note: Do not filter complete medium.
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Subculturing |
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 culture medium.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.05% (w/v) Trypsin- 0.53 mM EDTA solution containing 0.5% polyvinylpyrrolidone (PVP)
- Add 2.0 to 3.0 mL of Trypsin-EDTA-PVP solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually with 5 to 10 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 2.0 to 3.0 mL 0.1% Soybean Trypsin inhibitor and aspirate cells by gently pipetting.
- To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
- Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new coated culture vessels.
- Place culture vessels in incubators at 37°C.
Subcultivation ratio: 1:3 to 1:4
Medium Renewal: Every 2 to 3 days.
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.
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Cryopreservation |
Freeze medium: Leibovitz's L-15 medium with 2 mM L-glutamine and 10mM HEPES supplemented with 1% PVP, 10% fetal bovine serum and 7.5% 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: 10,12 D13S317: 11 D16S539: 9,11 D5S818: 11,12 D7S820: 9 THO1: 7 TPOX: 11 vWA: 16 |
Name of Depositor |
CC Harris |
Deposited As |
human |
Year of Origin |
1986 |
References |
Stoner GD, et al. Establishment and characterization of SV40 T-antigen immortalized human esophageal epithelial cells. Cancer Res. 51: 365-371, 1991. PubMed: 1703038
Milo GE, et al. Conversion of premalignant human cells to tumorigenic cells by methylmethane sulfonate and methylnitronitrosoguanidine. Cell Biol. Toxicol. 8: 193-205, 1992. PubMed: 1337307
Tolleson WH, et al. Apoptotic and anti-proliferative effects of fumonisin B1 in human keratinocytes, fibroblasts, esophageal epithelial cells and hepatoma cells. Carcinogenesis 17: 239-249, 1996. PubMed: 8625445
Tolleson WH, et al. The mycotoxin fumonisin induces apoptosis in cultured human cells and in livers and kidneys of rats. Adv. Exp. Med. Biol. 392: 237-250, 1996. PubMed: 8850621
Wan X, et al. Synthetic retinoid CD437 induces apoptosis of esophageal squamous HET-1A cells through the caspase-3-dependent pathway. Anticancer Res. 21: 2657-2663, 2001. PubMed: 11724335
Stoner GD, et al. Establishment and characterization of SV40 T-antigen immortalized human esophageal epithelial cells. Cancer Res. 51: 365-371, 1991. PubMed: 1703038
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