Summary
Surface coverage with autogeneous endothelial cells is effective in reducing thrombogenicity
of an artificial vascular graft, but procedure for obtaining the cells is invasive
for patients. The purpose of this study was to establish cultures of human endothelial
cells separated from a small piece of subcutaneous fat tissue. A piece of tissue weighing
about 10 mg was obtained from subcutaneous fat using a biopsy needle, and treated
with collagenase and dispase. Microvascular endothelial cells were selected and other
types of cells contaminating the cultures were eliminated by scraping with a needle
under a microscope. The yield of the cells was 8362 ± 4264/10 mg of subcutaneous fat
(n = 7). The cultures reached confluence in about 2 weeks. The cells were positive
for von Willebrand factor, P-selectin, and uptake of acetylated low density lipoprotein.
The cells produced 15.9 ± 3.3 ng/mg cell protein/h of 6-ketoprostaglandin F1α (n = 5) when stimulated with thrombin. Thrombin also stimulated the production of
platelet-activating factor: 7653 ± 4297 dpm/106 cells (n = 5). Endothelin-1 accumulation in the medium of unstimulated endothelial
cells was 0.54 ± 0.16 ng/mg cell protein/10 h (n = 8). As a preliminary experiment
for graft seeding, the cells were also cultured on pieces of a gelatin-coated Dacron
graft, and scanning electron microscopy revealed the surface coverage of the graft.
We herein described about successful culture of human microvascular endothelial cells
from subcutaneous fat tissue obtained using a biopsy needle. The cultured cells may
be applicable to a seeded vascular graft.
