Immortalized Human Corneal Endothelial Cells (IHCE) | T0577

Immortalized Human Corneal Endothelial Cells (IHCE) | T0577

Together with the Immortalized Human Corneal Keratinocytes (T0579) and the Immortalized Human Corneal Fibroblasts (T0578) 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

Organism:

Homo sapiens

Species:

Human

Source Organ:

Descemets′ membrane

Organ Type:

Eye

Growth Properties:

Adherent

Morphology:

Cobble-stone

Passage Number:

N/A

Population Doubling:

54 hours

Seeding Density:

20, 000 cells/cm10²; Split ratio of 1:2 to 1:3

Markers:

CX43, ZO-1, 9.3E

Donor Age:

N/A

Donor Gender:

Male

Donor Ethnicity:

N/A

Propagation:

The base medium for this cell line is Prigrow I . To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%, 5 mg/L human insulin (Z101065), 10 µg/ml human transferrin (Sigma; T8158), 3 ng/ml sodium selenite (Sigma; S5261), 10 nM hydrocortisone (Sigma; H0888), 10 nM β-estradiol (Sigma; E2758), 10 ng/ml human VEGF 165aa (Z100895), 10 ng/ml human EGF (Z100135), 10 ng/ml Heparin (Sigma; H3149), L-glutamine to a final concentration of 1%, and Penicillin/Streptomycin Solution to a final concentration of 1%.

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 CX43, ZO-1 and 9.3E; 3) FISH was used to examine the cytogenetic alterations in the immortalized cells.

Shipping Condition:

Dry Ice

Storage Condition:

liquid nitrogen or -180C