In vitro studies of human choroidal endothelial cells.
Vascular endothelial cells play an important role in progression or healing of various retinal or choroidal diseases and they have a broad organ specificity. We have grown and studied human choroidal endothelial cells (CECs) cultured in a collagen gel, where they form tube-like structures. CECs were differentially isolated from choroidal tissues and cultured to near homogeneity. The cells were embedded in a type I collagen gel in 24-well culture dishes and incubated with M199 medium containing 10% fetal calf serum. Gels were observed using phase contrast microscopy and were evaluated histologically by light and electron microscopy. The CECs were strongly positive for factor VIII-related antigen and actively ingested diI-acetylated LDL, indicating their endothelial nature. Ultrastructural analysis of the tube-like structures revealed a central lumen surrounded by cells joined apically by junctions and showing prominent pinocytotic activity, fenestrations and basement membrane formation; these features are typical of CECs in vivo. This preparation should be a useful tool to study the development of physiology and pathology of choroidal endothelium.
Doheny Eye Institute, Los Angeles, CA 90033, USA.
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T
Sakamoto
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H
Sakamoto
D R
DR
Hinton
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C
Spee
T
T
Ishibashi
S J
SJ
Ryan
D002232:Carbocyanines; D002469:Cell Separation; D002478:Cells, Cultured; D002829:Choroid / Q000098:blood supply*; D003001:Cloning, Molecular; D003094:Collagen; D003470:Culture Media; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D005169:Factor VIII / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D005782:Gels; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D018919:Neovascularization, Physiologic
C035005:3,3'-dihexadecylindocarbocyanine; D002232:Carbocyanines; D003470:Culture Media; D005456:Fluorescent Dyes; D005782:Gels; D008077:Lipoproteins, LDL; D005169:Factor VIII; D003094:Collagen