Immortalized Human Corneal Fibroblasts (IHCF)

Tissue repair and wound healing in the cornea is accomplished mainly by the corneal fibroblasts, and these cells are responsible for pathological process in the degenerative corneal disorders such as keratoconus. Established using the HPV16 E6/E7 genes, The Immortalized Human Corneal Fibroblasts (IHCF) not only retains its authentic cell biomarkers (CD36, ECM protein collagen I and S100A4), 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 Keratinocytes (T0579) 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: Spindle shaped

Population Doubling: 70 - 80 hours

Seeding Density: 10,000 - 20,000 cells/cm²

Markers: CD36, ECM protein collagen I, S100A4

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow III (TM003). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10% and Penicillin/streptomycin. 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. * abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

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 CD36, ECM protein collagen I, and S100A4; 3) FISH was used to examine the cytogenetic alterations in the immortalized cells.

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C