Het-1A (ATCC® CRL-2692)

Organism: Homo sapiens, human  /  Cell Type: epithelial; SV40 large T antigen transfected  /  Tissue: esophagus  /  Disease: normal

Permits and Restrictions

View Permits View Restrictions

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]
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 RefStoner 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.
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.

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.
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.

  1. Remove and discard culture medium.  
  2. Briefly rinse the cell layer with 0.05% (w/v) Trypsin- 0.53 mM EDTA solution containing 0.5% polyvinylpyrrolidone (PVP)  
  3. 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.
  4. Add 2.0 to 3.0 mL  0.1% Soybean Trypsin inhibitor and aspirate cells by gently pipetting.   
  5. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.  
  6. Discard supernatant and resuspend cells in fresh growth medium.  Add appropriate aliquots of cell suspension to new coated culture vessels.
  7.  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.  

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
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

Notice: Necessary PermitsPermits

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
Restrictions

The cells are distributed for research purposes only. The National Cancer Institute (NCI) releases the line subject to the following: 1.) The cells or their products must not be distributed to third parties. Neither the cell line nor products derived from it may be sold or used for commercial purposes. 2.) Any commercial requests or proposed commercial use of these cells must first be negotiated with the Office of Technology Transfer, NIH, 6011 Executive Blvd., Room 325, Rockville, MD 20852; Telephone (301) 496-7057; FAX (301) 402-0220

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