Human corneal epithelial primary cultures and cell lines with extended life span: in vitro model for ocular studies.
OBJECTIVE
To develop an in vitro model of human corneal epithelium that can be propagated in serum-free medium that is tissue specific, species specific, and continuously available.


METHODS
Primary explant cultures from human cadaver donor corneas were generated and subsequently infected with Adeno 12-SV40 (Ad12-SV40) hybrid virus or transfected with plasmid RSV-T.


RESULTS
Several lines of human corneal epithelial cells with extended life span were developed and characterized. Propagation of both primary cultures and lines with extended life span, upon collagen membranes at an air-liquid interface, promoted multilayering, more closely approximating the morphology observed in situ.


CONCLUSIONS
In vitro models, using primary cultures of corneal epithelium and lines of corneal epithelial cells with extended life span, retain a variety of phenotypic characteristics and may be used as an adjunct to ocular toxicology studies and as a tool to investigate corneal epithelial cell biology.
Kahn
C R
CR
Gillette Medical Evaluation Laboratories, Gaithersburg, MD 20879.
Young
E
E
Lee
I H
IH
Rhim
J S
JS
D000260:Adenoviruses, Human / Q000254:growth & development; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002460:Cell Line; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003315:Cornea / Q000166:cytology* / Q000382:microbiology; D016895:Culture Media, Serum-Free; D004847:Epithelial Cells; D004848:Epithelium / Q000382:microbiology; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007621:Karyotyping; D007633:Keratins / Q000378:metabolism; D008875:Middle Aged; D008954:Models, Biological; D013539:Simian virus 40 / Q000254:growth & development; D014162:Transfection; D014709:Vero Cells
D016895:Culture Media, Serum-Free; D007633:Keratins