[Cell culture and its application. Application of cultured vascular endothelial cells to the in vitro assay for the invasion of cancer cells through endothelium-matrix membrane system].
Usefulness of cultured human vascular endothelial cells for the in vitro assay of cancer invasion was reviewed. Selective isolation procedures for the vascular endothelial cells from the different types of vascular tubes were described. Human umbilical cord vein is one of the most useful sources for the isolation of fresh normal endothelial cells. The cells are isolated by perfusing trypsin solution through the umbilical cord vein and successively subcultivated up to 80 population doubling levels as in vitro life span. In case of human thoracic aorta, endothelial cells were removed from endothelium layer of aorta ring by a jet flow of collagenase solution through a needle. Purity of these cells as endothelial cells can be determined by indirect staining of the cytoplasma with anti-endothelin antibody. Usefulness of these cultured endothelial cells has been proved in the studies of regulation of endothelin biosynthesis, ageing of vascular cell, isolation of some new growth factors, interaction with other blood cells, etc. In addition to these studies, we consider that interaction of tumor cells with endothelial cells plays an important role in tumor metastasis. So we established in vitro invasion assay system through the interaction with endothelial cells to examine the invasion and adhesion of tumor cells. Human umbilical cord endothelial cells (HUVEC) were cultured on porous membranes coated with laminin (LN). HT1080 fibrosarcoma cells were seeded onto HUVEC, and HT 1080 cells passed through the membrane were counted. HUVEC were easily distinguished from tumor cells by specific staining of endothelial cells with UEA-1 lectin. Using scanning electron microscopy, we confirmed that HT1080 cells invaded between HUVEC. Roles of adhesion molecules induced by some cytokines on invasion of cancer cells through the endothelial cells-matrix membrane system were discussed.
Okada
T
T
National Chemical Labolatory for Industry.
Okuno
H
H
Mitsui
Y
Y
D002448:Cell Adhesion / Q000502:physiology; D002450:Cell Communication; D002462:Cell Membrane / Q000473:pathology; D002463:Cell Membrane Permeability; D002469:Cell Separation / Q000379:methods; D002471:Cell Transformation, Neoplastic / Q000473:pathology*; D002478:Cells, Cultured; D016232:Endothelins / Q000096:biosynthesis; D004730:Endothelium, Vascular / Q000378:metabolism / Q000473:pathology*; D005354:Fibrosarcoma / Q000473:pathology*; D006801:Humans; D009361:Neoplasm Invasiveness
D016232:Endothelins