Kinetics of the endocytotic pathway of Low Density Lipoprotein (LDL) in human endothelial cells line under shear stress: an in vitro confocal microscopy study.
We studied the effect of mechanical forces (shear stress) on the kinetics of internalization of native LDL and ox-LDL in endothelial cell line ECV304. This study was performed by using Confocal microscopy and FRET with two carbocyanine dyes, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiO) as the donor and 3,3'-dioctadecyloxacarbocyanine perchlorate (DiI) as the receptor. The cells were incubated with a culture medium containing either 10 microg/ml DiI-LDL or DiO-LDL in static conditions or subjected to a laminar flow under a Confocal Laser Scanning Microscope (SP2 Leica, Germany). The results showed: (1) the possibility to evaluate the kinetics of LDL endocytosis in living cells, (2) shear stress in comparison with control group more effectively enhanced LDL uptake, (3) ox-LDL (>50 microg/ml) >4 hours incubation was found to affect the cells as reflected by their detachment at low shear stress.
Traoré
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Groupe de Mécanique et Ingénierie Cellulaire et Tissulaire, UMR-CNRS 7563 LEMTA et IFR 111 Bioingénierie, Faculté de Médecine, 54500 Vandoeuvre lès Nancy, France. Mariama.Traore@medecine.uhp-nancy.fr
Sun
R J
RJ
Fawzi-Grancher
S
S
Dumas
D
D
Qing
X
X
Santus
R
R
Stoltz
J-F
JF
Muller
S
S
D002460:Cell Line; D004705:Endocytosis; D042783:Endothelial Cells / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes; D006801:Humans; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000378:metabolism*; D018613:Microscopy, Confocal; D013314:Stress, Mechanical
D005456:Fluorescent Dyes; D008077:Lipoproteins, LDL; C099252:oxidized low density lipoprotein