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Extracellular vesicles: a mirror of cardiovascular diseases?
Germany (bvmd) - Ruhr-Universitaet, Bochum
Cardiology Research lab of Molecular Cardiology of the St.Josef Hospital and Bergmannsheil, clinics of the Ruhr-University of Bochum, St.Josef-Hospital, Gudrunstr. 56, 44801 Bochum
Prof. Dr. med Andreas Muegge
Prof. Dr. rer. nat. Kornelia Jaquet (head of the research lab); Dr. rer nat. Diana Cimiotti (post-doc)
Type of Research Project
- Basic science
What is the background of the project?
Extracellular vesicles are released from all type of cells in healthy and diseased state. They build a complex communication network due to their different cargos (proteins, lipids, nucleic acids). Three main vesicle types according to their size can be differentiated: apoptotic bodies, microvesicles and exosomes. We isolate the vesicles from blood of patients with atrial fibrillation (AF), coronary heart disease (CHD) and chronic obstructive pulmonary disease (COPD) as well as from healthy probands, but also from cardiomyocytes and human umbilical vein endothelial cells (HUVEC). We further analyze the vesicle composition (apoptotic bodies, microvesicles, exosomes) and their cargos, especially protein and nucleic acid composition. Additionally we are interested in their effects on endothelial cells and cardiomyocytes. The aim of the project is the better understanding of cell-cell communication and to identify vesicles as new biomarkers for cardiac diseases. To isolate extracellular particles from blood platelet free plasma (PFP) is generated by centrifugation. Apoptotic vesicles and microvesicles are isolated stepwise by centrifugation, exosomes by using an exosome extraction kit. The different vesicles are characterized by binding fluorescence labeled annexin V to phosphatidylserine on the surface of the vesicles (characteristic for nearly all vesicles). The labeled vesicle fractions are then analyzed using flow cytometry (FACS=fluorescence activated cell sorting). To investigate the effects of the different vesicles on endothelial and cardiac muscle cells, the vesicles are incubated with either HUVEC or freshly isolated cardiomyocytes from adult rat ventricles and for example cytotoxicity or apoptosis is measured using specific cell-based assay kits. To analyze their cargos, proteins and nucleic acids, RNA and DNA are extracted and separated by electrophoresis. E.g. proteins may be identified by Western blotting and mass spectrometric analysis.
What is the aim of the project?
What techniques and methods are used?
What is the role of the student?
What are the tasks expected to be accomplished by the student?
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
The student should be able to communicate in English fluently. He should also be able to write in English. He/She should be able to work in a team and have at least very basic laboratory skills (calculate buffer ingredients, pipetting, centrifugation) Subjects passed: lab courses (biochemistry or/and molecular biology)
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students
- Yanez Mo M; et al. (2015) Biological properties of extracellular vesicles and their physiological functions. J. Extracell Vesicles 4; p27066
- Witwer KW et al (2013) Standardization of ample collection; isolation and analysis in extracellular vesicle research. J Extracell Ves Res 2;p 20360
- Sluijter; JPG et al. (2014) Microvesicles and exosomes for intracardiac communication. Caediovasc Res 102
- Shantsila E et al. (2014) Circulating microparticles: challenges and perspectives of flow cytometric assessment. Thromb. Haemos. 111; p1009 til 1114
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