Inhibition of vascular endothelial cell morphogenesis in cultures by limbal epithelial cells.
OBJECTIVE
To study the in vitro angiogenic activity of human conjunctival and limbal epithelial cells and conjunctival, limbal, and corneal fibroblasts in a three-cell-type coculture model.


METHODS
Human umbilical vein endothelial cells (EC) were cocultured with epithelial cells, fibroblasts, or epithelial cells and fibroblasts to test their effect on EC morphogenesis. Neutralizing antibodies to some known angiogenic factors were added to the culture to see whether the EC morphogenesis may be blocked by a particular antibody.


RESULTS
Conjunctival and limbal epithelial cells exhibited very little or no stimulatory effect on EC tube formation when examined in an EC- epithelial cell coculture system. In contrast, conjunctival, limbal, and corneal fibroblasts all promoted EC morphogenesis when examined under the same culture conditions. Fibroblast-induced EC morphogenesis was inhibited by addition of anti-vascular endothelial growth factor (VEGF) and/or anti-basic fibroblast growth factor (bFGF) antibodies to the culture medium. In the three-cell-type coculture system consisting of ECs, fibroblasts, and epithelial cells, limbal epithelial cells (but not conjunctival epithelial cells) exhibited a strong inhibitory effect on fibroblast-induced EC tube formation.


CONCLUSIONS
The proangiogenic activity of ocular surface fibroblasts is probably mediated through a paracrine mechanism by VEGF and bFGF. Limbal epithelial cells, but not conjunctival epithelial cells, inhibit fibroblast-stimulated angiogenesis.
Department of Ophthalmology, Change Gung Memorial Hospital, Tao-Yuan, Taiwan, Republic of China.
D H
DH
Ma
R J
RJ
Tsai
W K
WK
Chu
C H
CH
Kao
J K
JK
Chen
D000293:Adolescent; D000328:Adult; D000368:Aged; D002454:Cell Differentiation; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D018920:Coculture Techniques; D003228:Conjunctiva / Q000166:cytology; D003315:Cornea / Q000166:cytology; D016228:Endothelial Growth Factors / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology*; D004847:Epithelial Cells / Q000502:physiology*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005347:Fibroblasts / Q000502:physiology; D006801:Humans; D016850:Limbus Corneae / Q000166:cytology; D008222:Lymphokines / Q000494:pharmacology; D008875:Middle Aged; D009024:Morphogenesis; D018919:Neovascularization, Physiologic / Q000187:drug effects; D019899:Paracrine Communication; D014471:Umbilical Veins / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D016228:Endothelial Growth Factors; D008222:Lymphokines; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D016222:Fibroblast Growth Factor 2