Location & dates EMBL Heidelberg, Germany 26 - 28 Nov 2014
Deadlines Application closed

Programme

Day 1 - Wednesday 26 November
TimeSpeakerTitle
  General Introduction
9:00 19:45

Welcome
Yvonne Yeboah - EMBL and An Hendrix - Ghent University

 

Guillaume Van Niel, Olivier De Wever - Ghent University

Exosome biogenesis and function
 

EXOSOME PURIFICATION: obstacles and opportunities in the field

Coffee break  to get to know each other. 15 participants present their research focus and/or topic related problems that can be solved during the course (5 min/participant).

Dimitri Aubert - Izon

Single-step isolation of extracellular vesicles by Size Exclusion Chromatography

Ketil Winther Pedersen - Life Technologies

Cell culture exosome isolation and analysis: Generic enrichment (precipitation) and specific isolation (magnetic beads)

Davide Zocco - Hansabiomed

Total exosome RNA extraction following specific immunocapture from biological fluids

Lunch

Martin Schlumpberger - Qiagen

Membrane affinity spin columns to efficiently isolate RNA from exosomes and other extracellular vesicles (ExoRNeasy)

An Hendrix - Ghent University and Esther Nolte-‘t Hoen - University of Utrecht

Density gradient ultracentrifugation (Optiprep and Sucrose)

Tamas Matusek - CNRS

The role of ESCRT in the release of Hh containing EVs and technical challenges to isolates EVs from limited amounts of animal fluids

Qiagen

Column-based capture (ExoRNeasy):
Coffee break

Practical:  Demonstration of exosome isolation by density gradient ultracentrifugation (sucrose and optiprep) and commercial kits from Izon, Hansabiomed, Life Technologies and Qiagen

Dinner
Day 2 - Thursday 27 November
TimeSpeakerTitle
9:00- 19:15

EXOSOME CHARACTERIZATION: How to quantify and assess the purity of isolated exosome fractions and discuss obstacles and opportunities in the field

Guillaume Van Niel - Institut Curie
Exosome characterization by immunoblot and transmission electron microscopy
Marca Wauben - University of Utrecht Quantitative and qualitative analysis by flow cytometry

Coffee break

Vincent Hyenne - INSERM U1109

Combination of intra-vital imaging and correlated electron microscopy to monitor exosomes in a living mouse

Jörg Uhlig - Malvern Nanoparticle Tracking Analysis: Nanosight and qNano
Dimitri Aubert - Izon qNano and Tunable Resistive Pulse Sensing
Lunch
Practical (1): Continue with exosome isolation from day 1
Coffee break
Practical (2): Demonstration of exosome characterization by transmission electron microscopy, Nanoparticle Tracking Analysis and Tunable Resistive Pulse Sensing
Day 3 - Friday 28 November
TimeSpeakerTitle
9:00-16:30

EXOSOME COMPOSITION: obstacles and opportunities in the field

Pieter Mestdagh  - Ghent University Transcriptomics (1)

Jo Vandesompele - Ghent University

Transcriptome validation by Q-PCR

Coffee break

Esther Nolte-‘t Hoen - University of Utrecht Analysis of extracellular vesicle RNA: what to expect and consider

Michael Pfaffl - Technische Universitaet Muenchen

Exosome isolation and holistic expression profiling using RNA-Seq and RT-qPCR
Lunch
Andrea Németh - Semmelweis University

Label-free GCI-technique to measure exosome adhesion on extracellular matrices

Johan Swinnen - University of Leuven Lipidomics
An Hendrix - Ghent University Proteomics
Coffee break
Olivier De Wever - Ghent University Summary of take home messages
Course Ends