Disease:

Normal

Species:

Homo sapiens, human

Tissue:

Colon

Morphology:

Epithelial; although FHC cells exhibit epithelial morphology, cytoplasmic keratins were not detected.

Properties:

Adherent

Patient:

13 weeks gestation.

Medium:

A 1:1 mixture of DMEM:F12 Medium.To make the complete growth medium, add the following components to the base medium: extra 10 mM HEPES (for a final conc. of 25 mM); 10 ng/ml cholera toxin; 0.005 mg/ml insulin; 0.005 mg/ml transferrin; 100 ng/ml hydrocortisone; 20 ng/mL human recombinant EGF; fetal bovine serum 10% (final conc.)

Subculture:

Remove medium, and rinse with 0.05% trypsin, 0.53 mM EDTA solution. Remove the solution and add an additional 1 to 2 mL of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37°C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks. Culture at 5% CO2, 37°C. A subcultivation ratio of 1:2 to 1:4 is recommended every 10-15 days. Renew medium once every 3 to 4 days depending on cell density.

Freezing Medium:

Complete growth medium supplemented with an extra 50% FBS and 10% (v/v) DMSO