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HIEC-6 (ATCC® CRL-3266)

Organism: Homo sapiens, human  /  Cell Type: epithelium  /  Tissue: small intestine  /  Disease: normal

Permits and Restrictions

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Organism Homo sapiens, human
Tissue small intestine
Cell Type epithelium
Product Format frozen 1.0 mL
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 1

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 17 to 19 week gestation
Applications

Investigation of the regulation of normal intestinal cell proliferation, survival, stemness, as well as studies of the molecular mechanisms leading to lineage-specific differentiation. Also used for the study of the effects of hormones, growth factors, extracellular matrix molecules, drug metabolism and carcinogenic factors on small intestinal functions.

Frequently used as normal control cells for studies dealing with colorectal cancer cell lines.

Storage Conditions liquid nitrogen vapor phase
Images CRL-3266 Cell Micrograph
Derivation HIEC-6 cells were isolated by thermolysin treatment of a human fetal small intestine.
Antigen Expression cytokeratin-8 +, cytokeratin-18 +, cytokeratin-21 +, mouse intestinal mucosa (MIM-1/39) +, aminopeptidase N (APN) +, dipeptidyl-peptidase IV (DPPIV) +
Tumorigenic no
Comments This is a continuously growing human intestinal epithelial cell line which expresses epithelial cytokeratins and functional markers for intestinal crypt cells. They exhibit typical intestinal crypt cell proliferative and undifferentiated characteristics.
Complete Growth Medium The base medium for this cell line is OptiMEM 1 Reduced Serum Medium (Gibco Catalog No. 31985). To make the complete growth medium, add the following components to the base medium:
  • 20 mM HEPES
  • 10 mM GlutaMAX (Gibco Catalog No. 35050)
  • 10 ng/mL Epidermal Growth Factor (EGF)
  • fetal bovine serum (FBS) to a final concentration of 4%

Subculturing Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) (ATCC® 30-2200) or 0.25% (w/v) Trypsin-0.53 mM EDTA solution (ATCC® 30-2101) to remove all traces of serum that contains trypsin inhibitor.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended 

Medium Renewal: Every 2 to 3 days

Cryopreservation

Freeze medium: Complete Growth Medium 95%; DMSO (ATCC 4-X), 5%. Storage temperature: liquid nitrogen vapor phase

Culture Conditions Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Volume 1.0 mL
STR Profile Amelogenin: X
CSF1PO: 11,12
D13S317: 11,14
D16S539: 12,13
D5S818: 11,13
D7S820: 8,11
TH01: 6,9
TPOX: 8,11
vWA: 14,15
Population Doubling Time 96 hours
Name of Depositor JF Beaulieu
Year of Origin 1995
References

Perreault N, et al. Use of the dissociating enzyme thermolysin to generate viable human normal intestinal epithelial cell cultures. Exp Cell Res 224(2): 354-364, 1996. PubMed: 8612712

Pageot LP, et al. Human cell models to study small intestinal functions: recapitulation of the crypt-villus axis. Microsc Res Tech 49(4): 394-406, 2000. PubMed: 10820523

Guezguez A, et al. Modulation of stemness in a human normal intestinal epithelial crypt cell line by activation of the WNT signaling pathway. Exp Cell Res 322: 355-364, 2014. PubMed: 24534551

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
Other Documentation
FAQ's
  1. Maximum number of passages for CRL-3266
    ATCC has not performed a longevity study on these cells but per information received from the depositor, the maximum number of passages is 28.
    Date Updated: 5/26/2017
References

Perreault N, et al. Use of the dissociating enzyme thermolysin to generate viable human normal intestinal epithelial cell cultures. Exp Cell Res 224(2): 354-364, 1996. PubMed: 8612712

Pageot LP, et al. Human cell models to study small intestinal functions: recapitulation of the crypt-villus axis. Microsc Res Tech 49(4): 394-406, 2000. PubMed: 10820523

Guezguez A, et al. Modulation of stemness in a human normal intestinal epithelial crypt cell line by activation of the WNT signaling pathway. Exp Cell Res 322: 355-364, 2014. PubMed: 24534551