Immortalized Human Corneal Keratinocytes (IHCK)

Corneal Keratinocytes play a major role in maintaining the balance of stromal substances and preserving the corneal transparency. Established using the HPV16 E6/E7 genes, The Immortalized Human Corneal Keratinocytes (IHCK) not only retains its authentic cell biomarkers (CK12, CK19), it also displays fairly disomic cytogenetic status at chromosomes 1, 8, 10 and 18, which is an indication of normal diploid chromosomal status.

Together with the Immortalized Human Corneal Stromal Fibroblasts (T0578) and the Immortalized Human Corneal Endothelial cells (T0577) isolated from the same donor and immortalized using the same platform, this uniform model system presents a valuable tool for studies involving cornea homeostasis and ophthalmologic diseases.

Immortalization Method: Serial passaging and transduction with retroviruses carrying human papillomavirus type 16 E6/E7 gene

BioSafety Level: II

Species: Human

Source Organ: corneoscleral rim

Growth Properties: Adherent

Morphology: Polygonal

Population Doubling: 24 hours

Seeding Density: 30,000 - 50,000 cells/cm²

Markers: CK19, CK12, CD36, Stromal Fibrillar Collagen Type 1, S100A4

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow X series medium available at abm, Cat. No. TM0579. The cells carry a neomycin resistant marker, thus may culture the cells with 20 µg/ml G418.
Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.

Quality Control: 1) RT-PCR was used to evaluate the transgene expression of HPV16 E6/E7; 2) Indirect immunofluorescence was performed to detect the cell specific marker expression CK12, CK19 and ABCG2; 3) FISH was used to examine the cytogenetic alterations in the immortalized cells.

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C