Fluorescent probe-labeled lipid microsphere uptake by human endothelial cells: a flow cytometric study.
Lipid microspheres have been used as carriers of drugs such as prostaglandin E1 (lipo-PGE1) and corticosteroid (liposteroid). Lipo-PGE1 is used for the treatment of chronic arterial occlusive diseases because its activity is far greater than that of free PGE1 in vivo. To verify the fact that the drug carriers, lipid microspheres, are preferentially taken up by endothelial cells, we labeled lipid microspheres with a fluorescent probe, DiI (DiI-LM), and observed them in some in vitro models. Stoichiometric fluorescence was obtainable, and the fluorescence was stable between pH 3.3 and pH 8.9. Human umbilical vein endothelial cells and cells of a human endothelial cell line, ECV304, showed increased uptake of DiI-LM, 81% and 61%, respectively. In contrast, uptakes were less than 7% in human skin fibroblasts, 3T3 cells, and human neutrophils. Prominent perinuclear fluorescence was also observed in endothelial cells by fluorescence microscopy. DiI-LM and flow cytometric analysis will be useful for studies to elucidate the precise mechanism of the selective accumulation of lipid microspheres by cells in blood vessel walls.
Suzuki
K
K
Internal Medicine I, National Defense Medical College, Saitama, Japan.
Takahashi
K
K
Matsuki
Y
Y
Kawakami
M
M
Kawaguchi
Y
Y
Hidaka
T
T
Sekiyama
Y
Y
Mizukami
Y
Y
Kawagoe
M
M
Nakamura
H
H
D002232:Carbocyanines; D002448:Cell Adhesion; D002460:Cell Line; D004655:Emulsions; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D050356:Lipid Metabolism; D008863:Microspheres; D009504:Neutrophils / Q000378:metabolism; D012867:Skin / Q000166:cytology; D014280:Triglycerides / Q000378:metabolism
D002232:Carbocyanines; D004655:Emulsions; D005456:Fluorescent Dyes; D014280:Triglycerides; C024286:3,3'-dioctadecylindocarbocyanine