Mobirise

PROJECT 3 

Vascular smooth muscle cell phenotypic switching.  

PROJECT DESCRIPTION

Objectives: 

Test the hypothesis that microcalcifications are a consequence from phenotypic switching of vascular smooth muscle cells.


Methodologies: 

1. Primary VSMCs will be isolated from human aortic tissue. VSMCs will be cultured in the presence of growth factors (i.e PDGF) to induce synthetic VSMCs or TGFb or heparin to obtain contractile VSMCs. Phenotypic diversity will be measured and validated by qPCR and western blotting and compared with data from ESR7. Results will be compared to contractile and synthetic pig VSMCs, an accepted model for phenotypic diversity of VSMCs.


2. As no markers for synthetic VSMCs are present, we will screen for novel markers indicative for synthetic VSMCs. Utilising a miniaturised cell-based platform to analyse differences in proliferation, migration and invasion (xCELLigence®) and cell functions by multiplex fluorescence analyses using established and newly developed cell-function-reporters. Novel markers will be tested in ESR6 and ESR12 (mouse models of microcalcification).


3. In vivo, we will monitor VSMC phenotype in apoE:sm22ahDTr and in smoothelin driven cherry reporter mice, available in our lab in order to address the role of VSMC apoptosis in microcalcification. These models will also be utilised in ESR11 to detect upstream pathways of microcalcification. In vivo effects of CKD and DMII, (and impact of pharma/nutraceutical treatment) on VSMC phenotypic switching will be assessed in the smoothelin reporter.

Host institution

CARIM Maastricht University (the Netherlands) 

Dr Leon Schurgers