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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   #
1   Department of Cardiothoracic and Vascular Surgery
,
Michael Torzewski   #
2   Institute of Clinical Chemistry and Laboratory Medicine, University of Mainz, Mainz, Germany
,
Manfred Dahm
1   Department of Cardiothoracic and Vascular Surgery
,
Viola Ochsenhirt
2   Institute of Clinical Chemistry and Laboratory Medicine, University of Mainz, Mainz, Germany
,
Hans-Anton Lehr
3   Institute Universitaire de Pathologie, Centre Hospitalier Universitaire Vaudois (CHUV), Lausanne, Switzerland
,
Karl J. Lackner
2   Institute of Clinical Chemistry and Laboratory Medicine, University of Mainz, Mainz, Germany
,
Christian-Friedrich Vahl
1   Department of Cardiothoracic and Vascular Surgery
› Author AffiliationsFinancial support: This work was supported in part by the University of Mainz (MAIFOR A11 to B.D.) and the German Research Foundation (SFB 432, Z3 to H.A.L.).
Further Information

Publication History

Received 24 May 2005

Accepted after revision 28 October 2005

Publication Date:
28 November 2017 (online)

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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

# Both authors contributed equally to this work.


 

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