Kinetics of the endocytotic pathway of Low Density Lipoprotein (LDL) in human endothelial cells line under shear stress: An in vitro confocal microscopy study
Issue title: Selected Papers from the First Seminar on “Medical Engineering and Therapy”, 27–28 April 2004, Nancy, France
Affiliations: Groupe de Mécanique et Ingénierie Cellulaire et Tissulaire, UMR-CNRS 7563 LEMTA et IFR 111 Bioingénierie, Faculté de Médecine, 54500 Vandoeuvre lès Nancy, France | Research Institute of Clinical Medical Sciences, China–Japan Friendship Hospital, Beijing 100029, China | Muséum National d'Histoire Naturelle, Laboratoire de Photobiologie, 43 rue Cuvier 75231, Paris cedex 05, France
Note: [] Corresponding author: Mariama Traore, Groupe de Mécanique et Ingénierie Cellulaire et Tissulaire, UMR-CNRS 7563 (LEMTA), Faculté de Médecine BP: 184, 54500 Vandoeuvre lès Nancy, France. Tel.: +33 0 3 83 68 34 56; Fax: +33 0 3 83 68 34 59; E-mail: Mariama.Traore@medecine.uhp-nancy.fr.
Note: [] Present address: Department of Life Sciences, National Natural Science Foundation of China, 100085 Beijing, PR of China.
Abstract: 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 μg/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 μg/ml) >4 hours incubation was found to affect the cells as reflected by their detachment at low shear stress.
Keywords: Atherosclerosis, low density lipoproteins, shear stress, confocal microscopy, kinetics
