Objective
Fibroblasts will be isolated and cultured from different rat organs by the group in Geneva. Fibroblasts will be cultured from the following organs: dermis, subcutaneous tissue, striated muscle, heart, lungs, tendon and intestinal wall. The cells will be brought up to the fifth passage and characterised as far as their replicative activity and expression of cytoskeletal and contractile proteins are concerned. In particular, the percentage of cells expressing a smooth muscle actin, the expression of a smooth muscle actin by Western blots and of a smooth muscle actin mRNA by Northern blots will be measured. When the cells are at the fifth passage, they will be also characterised by their production of collagen type 1 and type 3, fibronectin, lisyl oxidase and proteoglycans. This part, analysing extracellular matrix compound production, will be done in Lyon.
Motility of the different fibroblasts will be studied in Geneva and Moscow by different techniques. In Moscow, characteristics such as speed, directionality and especially pseudo-podial activity will be quantified. Moreover, parameters characterising cell shape and spreading, in particular, polarity of the cell outline will be evaluated. Finally, intracellular particles movement including average velocity and distribution of the particles in the cytoplasm will be measured.
The experiments will be done in Geneva and in Moscow. The isolation and characterisation of cytoskeletal features in fibroblasts of different organs will be performed mainly during the first year of the contract. Then extracellular matrix proteins will be studied in Lyon and cell motility will be studied in Geneva and in Moscow. Polarisation, cell shape spreading and particle movement will be all studied in Moscow during the second and third years of the project.
Topic(s)
Call for proposal
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1211 Genève
Switzerland