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Methods available for cooperations

 

Please find below a summary of methods and techniques available for cooperations. For inquiries please contact Prof. Dr. Thomas Thum.

 

 

MicroRNA based techniques

  • microRNA (miRNA, miR) profiling in cells, tissues and different body fluids (e.g. plasma, serum, urine) by robotics-assisted quantitative Real-time PCR (qRT-PCR)
  • miRNA library screening applying a high-throughput transfection setup with an automated pipet robot system (functional miRNA screening)
  • Identification of Argonaute 2 (Ago2) bound mRNAs to analyze miRNA target genes by RNA immunoprecipitation (RIP) and subsequent genechip analysis (RIP-Chip)
  • Valdiation of miRNA target genes by luciferase reporter assays (luciferase assay miRNA target)
  • miRNA promoter analysis by luciferase reporter assays (luciferase assay miRNA promotor)

 

Cell culture and other molecular biology techniques

  • Testing of endothelial cell (EC) function: capillary formation/sprouting capacity (tube assay, spheroid assay) and migratory ability (Boyden-chamber migration assay)
  • Cell fractionation from adult mouse hearts
  • FACS (Guava cytometer- Millipore):
    • Surface marker expressions on different cells eg. platelets (CD-61, CD-41a),leukocytes (CD-45), endothelial cells (CD-31)
    • Evaluation of Cy-3 labelled pre-miRNA transfection efficiency
    • Apoptosis assays
  • Immunofluorescence microscopy (Nikon)
  • Magnetic cell sorting using Gentle MACS/ separation columns (Miltenyi Biotech)

 

Cardiovascular phenotyping

Standardized, high quality, state-of-the-art techniques are necessary to analyze cardiovascular function in mice in-vivo. We provide a wide range of methodologies that include:

  • Assessment of blood pressure, left/right ventricular function using Millar PV Catheter System (MPVS) at baseline and at endpoint of disease models. The MPVS allows simultaneous and continuous high-fidelity monitoring of left or right ventricular pressure and relative volume in the intact heart of anesthetized animals. Intraventricular pressure and volume signals generated in real time are the characteristic pressure-volume (P-V) loops, which are used to determine load-dependent and -independent parameters of cardiac contractile function in normal and disease conditions.
  • Assessment of cardiovascular function in genetically modified mice
  • Evaluation of treatment effects (acute or chronic, pharmacologic or biologic therapeutics)

Evaluation of stress models

Our understanding of cardiovascular disease development and our ability to develop preventive measures and treatments depend on suitable animal models that replicate human disease processes. We use well-characterized acute and chronic mouse models of various cardiac diseases.

Cardiac disease models:

  • MI: Myocardial infarction: LAD ligation without reperfusion
  • I/R: Myocardial infarction: LAD ligation with reperfusion
  • TAC: transverse aortic constriction, model of cardiac hypertrophy and remodelling
  • AII: Angiotensin-II minipump-infusion, model of hypertension and cardiac remodelling

Vascular models:

  • Perivascular carotid artery electric injury: model of vascular remodelling

In-vivo models of angiogenesis:

Subcutaneous implantation of angioreactors or matrigel plugs in mice and subsequent evaluation of angiogenesis. These models provide a standardized platform for reproducible and quantifiable in-vivo angiogenesis assays:

  • Angioreactors: Cultrex Directed In Vivo Angiogenesis Assay
  • Matrigel Plug Assay