In vitro studies of human choroidal endothelial cells

Curr Eye Res. 1995 Aug;14(8):621-7. doi: 10.3109/02713689508998488.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Carbocyanines
  • Cell Separation
  • Cells, Cultured
  • Choroid / blood supply*
  • Cloning, Molecular
  • Collagen
  • Culture Media
  • Endothelium, Vascular / cytology*
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / ultrastructure
  • Factor VIII / metabolism
  • Fluorescent Antibody Technique
  • Fluorescent Dyes
  • Gels
  • Humans
  • Lipoproteins, LDL / metabolism
  • Microscopy, Electron
  • Microscopy, Phase-Contrast
  • Neovascularization, Physiologic

Substances

  • 3,3'-dihexadecylindocarbocyanine
  • Carbocyanines
  • Culture Media
  • Fluorescent Dyes
  • Gels
  • Lipoproteins, LDL
  • Factor VIII
  • Collagen