Ex vivo technologies

Genetic engineering is the manipulation of an organism’s genes using biotech methodologies and it includes several applications that are widely adopted to study cardiac biology. In the past years, important advances have been achieved in the field (viral vector-mediated gene transfer, CRISPR cas9 gene editing etc) making these technologies rapid, efficient and more affordable. These methods are now used to answer specific research questions and they are applied to both in vitro and in vivo research models. The group is testing these novel technologies in a regenerative medicine perspective.

Working closely with the other research groups within Prof Thomas Thum's laboratory, and their great expertise in non-coding RNAs (ncRNAs), the group research interest is mainly focused on the role of ncRNAS in regeneration and the evaluation of both structural and functional parameters which are affected by the therapeutic treatments. The group uses several in vitro and ex vivo models, with a particular focus on living myocardial slice. In a translational perspective, human myocardial slices are also extensively used. The multicellular environment and preserved functionality during in vitro culture make living myocardial slices the ideal model to develop and test new therapeutic strategies.

Group members:

  • Perbellini Filippo, PhD, Group Leader (see CV)
  • Naissam Abbas, MD, PhD student
  • Mitzka Saskia, Technician
  • Waleczek Florian Johann Gustav, medical student (StrucMed)
  • Haas Jonas, medical student (StrucMed)

 

Methodology:

  • Myocardia slice preparation and in vitro culture
  • AAV-mediated gene expression (gene therapy)
  • Functional analysis (contractility measurements, calcium transient recordings etc)
  • Cardiac cell isolation (cardiac fibroblasts, endothelial cells, cardiomyocytes, macrophages etc)
  • In vitro and ex vivo imaging
  • Optical clearing
  • FACS assays
  • Biochemical assays (metabolic activity, live/dead etc)

 

Projects:

  1.  Investigation of resident cardiac macrophages in living ventricular myocardial slices and their manipulation using novel molecular tools.
  2. Genetic manipulation of living myocardial slices for functional studies of non-coding RNA pathways
  3. Investigating the regenerative potential of microRNA and non-coding RNA for translational medicine