Immortalized Human Colonic Epithelial Cells (HCEC-1CT)

Description: Immortalized Human Colonic Epithelial Cells (HCEC-1CT) is a human colonic epithelial cell (HCEC) line isolated from healthy adult patient biopsy and immortalized by transduction with the cyclin-dependent kinase 4 (Cdk4) and human telomerase reverse transcriptase (hTERT). HCEC-1CT cells express stem cell markers, such as LGR5, BMI1, CD29, and CD44, and retain multilineage epithelial differentiation capability. Thus, HCEC-1CT cells are capable of differentiating into various epithelial lineages of the colon and forming self-organizing cysts in 3D organotypic culture in Matrigel. During differentiation, HCEC-1CT cells express colon epithelial cell-specific markers such as A33 and mucin 1. Compared to most HCEC cell lines of malignant origin, HCEC-1CT is a valuable cytogenetically normal and nontumorigenic cell line for studying colon stem cell biology, differentiation as well as the initiation and progression of colonic diseases, such as colorectal cancer.

Immortalization Method: Immortalized with retroviral Cdk4 and Human telomerase reverse transcriptase (hTERT)

BioSafety Level: II

Species: Human

Source Organ: Colon

Growth Properties: Adherent

Morphology: Compact cuboidal to spindle-shaped cells

Population Doubling: 18 - 24 hours

Seeding Density: N/A

Markers: LGR5, BMI1, CD29, and CD44
In proliferation: vimentin, α-smooth muscle actin
Upon growth arrest: cytokeratin 18, zonula occludens-1, mucins-1 and -2, antigen A33, and dipeptidyl peptidase 4

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is a mix of Prigrow III medium and Prigrow I medium, in 4:1 ratio respectively. Both are available at abm, Cat. No. TM003; abm, Cat. No. TM001. To make the complete growth medium, add the following components to the base medium: cosmic calf serum to a final concentration of 2%, Recombinant Human EGF (Z100135) to a final concentration of 25 ng/mL, hydrocortisone (Sigma) to a final concentration of 1 µg/mL, Insulin Solution (TM053) to a final concentration of 10 µg/mL, Human Holo Transferrin (G599) to a final concentration of 2 µg/mL, sodium selenite (Sigma) to a final concentration of 5 nM, and Gentamicin (G272) to a final concentration of 50 µg/mL.
Change media every 2-3 days.
Carbon dioxide (CO₂): 7%, Temperature: +37.0°C.
* Do not use heat-inactivated serum for cell culture unless specified otherwise.

Quality Control: 1) Western blot; 2) Immunocytochemistry; 3) Transmission electron microscopy; 4) Karyotype and DNA sequence analysis

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C