Journal of Lipid Research, Vol 24, 105-112, Copyright © 1983 by Lipid Research, Inc.
Isolation and characterization of endothelial cells from the epididymal fat pad of the rat
P Bjorntorp, GK Hansson, L Jonasson, P Pettersson and G Sypniewska
Endothelial cells from rat epididymal fat pad capillaries were isolated
from rats immediately after weaning. The cells were obtained after an
initial brief incubation with collagenase under conditions of minimal
breakage of cells. Adipocytes were removed by flotation and endothelial
cells were then obtained as cell aggregates by fractional filtration
procedures whereby intact tissue as well as free cells were removed. These
aggregates were then dispersed and cultured in supplemented medium 199
whereby a monolayer of cells with a growth pattern, numerous pinocytotic
vesicles, and intercellular junctions typical of endothelial cells were
obtained. Minor contaminations of precursor cells to adipocytes were absent
after one subculture. Here greater than 95% of the cells showed the
presence of Factor VIII. Further subcultures produced nonhomogenous cells
and decreasing rates of replication. The endothelial cells showed a very
low rate of triglyceride synthesis and release, and collected no visible
lipid upon prolonged cultures in the presence of an abundance of
triglyceride substrate. They bound lipoprotein lipase from rat adipocytes,
whereby the lipase was stabilized. This binding was released by heparin,
and the cells did not synthesize the enzyme.
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