A novel in vitro model for the study of plaque development in atherosclerosis

Bernhard Dorweiler; Michael Torzewski; Manfred Dahm; Viola Ochsenhirt; Hans-Anton Lehr; Karl J. Lackner; Christian-Friedrich Vahl

doi:10.1160/TH05-05-0357

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2006; 95(01): 182-189
DOI: 10.1160/TH05-05-0357

New Technologies, Diagnostic Tools and Drugs

Schattauer GmbH

A novel in vitro model for the study of plaque development in atherosclerosis

Bernhard Dorweiler^#

¹Department of Cardiothoracic and Vascular Surgery

,

Michael Torzewski^#

²Institute of Clinical Chemistry and Laboratory Medicine, University of Mainz, Mainz, Germany

,

Manfred Dahm

¹Department of Cardiothoracic and Vascular Surgery

,

Viola Ochsenhirt

²Institute of Clinical Chemistry and Laboratory Medicine, University of Mainz, Mainz, Germany

,

Hans-Anton Lehr

³Institute Universitaire de Pathologie, Centre Hospitalier Universitaire Vaudois (CHUV), Lausanne, Switzerland

,

Karl J. Lackner

²Institute of Clinical Chemistry and Laboratory Medicine, University of Mainz, Mainz, Germany

,

Christian-Friedrich Vahl

¹Department of Cardiothoracic and Vascular Surgery

› Institutsangaben

Abstract

Summary

For the study of atherogenesis in vitro, coculture systems have been devised, in which two or more cell types can be cultured in close contact to each other. Herein, we describe a novel in vitro model that aims at the simulation of the morphology ofa normal muscular artery allowing for the study of the initial events in atherosclerosis. Usinga modified fibrin gel as a scaffold for the coculture of endothelial cells (ECs) and smooth muscle cells (SMCs), we generated an autologous in vitro model with a multilayer growth of SMCs (intima-like structure) covered by an endothelium. The production of extracellular matrix (ECM) could be visualized histologically and verified by (i) ascorbic-acid dependent secretion of procollagenI into the supernatant and (ii) deposition of collagens I and III as well as laminin in the gel as assessed by immunohistochemistry. By BrdU-incorporation and Ki67 expression, the SMCs exhibited minimal proliferative activity, even when the culture period was extended to6 weeks. Lipoprotein insudation was investigated under simulated hypo-, normo-and hypercholesterolemic conditions through addition of 0.5, 1 or2 mg/mL LDL to the medium with subsequent time and dose dependend insudation of LDL. When human monocytes were added to the culture medium, infiltration and foam cell formation of macrophages and SMCs as well as expression of interleukin-8 (IL-8) was demonstrated. The in vitro model of the human vascular wall described herein appears to be suitable for the study of pivotal events in atherosclerotic plaque development. The applicability for long-term culture, the ability to study cell-matrix interactions and the opportunities for histomorphological and immunohistochemical examinations represent additional advantages of this model.

Keywords

In vitro model - endothelium - atherosclerosis - lipoprotein - foam cell

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Referenzen

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