HIEC-6
CRL-3266 ™
HIEC-6 is an epithelial cell that was isolated from the small intestine of a patient.
This cell line was deposited by JF Beaulieu.
Product category
Human cells
Organism
Homo sapiens, human
Cell type
epithelial cell
Morphology
epithelial-like
Tissue
Small intestine
Disease
Normal
Applications
3D cell culture
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogenSpecific 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.Volume
1.0 mL
Growth properties
Adherent
Derivation
HIEC-6 cells were isolated by thermolysin treatment of a human fetal small intestine.
Age
17 to 19 weeks gestation
Tumorigenic
No
Antigen expression
Cytokeratin-8 +, cytokeratin-18 +, cytokeratin-21 +, mouse intestinal mucosa (MIM-1/39) +, aminopeptidase N (APN) +, dipeptidyl-peptidase IV (DPPIV) +
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.Unpacking and storage instructions
Check all containers for leakage or breakage.
Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below -130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete 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%
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure
To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt.
If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.
Thaw the vial by gentle agitation in a 37°C water bath.
To reduce the possibility of contamination, keep the O-ring and cap out of the water.
Thawing should be rapid (approximately 2 minutes).
Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol.
All of the operations from this point on should be carried out under strict aseptic conditions.
Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium. and spin at approximately 150-400 x g for 8 to 12 minutes.
Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio).
It is important to avoid excessive alkalinity of the medium during recovery of the cells.
It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). pH (7.0 to 7.6).
Incubate the culture at 37°C in a suitable incubator.
A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.
Subculturing procedure
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
Remove and discard culture medium.
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.
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.
Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
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
Reagents for cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)
Population doubling time
Approximately 96 hrs
STR profiling
Amelogenin: X
CSF1PO: 11,12
D13S317: 11,14
D16S539: 12,13
D5S818: 11,13
D7S820: 8,11
THO1: 6,9
TPOX: 8,11
vWA: 14,15
Depositors
JF Beaulieu
Year of origin
1995