Visualization of osteoblast differentiation by means of luciferase-reporter constructs
Thursday 16 April 2009
Project
One of the pathological features characterizing MM is osteolysis of the bone, resulting from an impaired balance between bone formation by the osteoblasts and bone resorption by the osteoclasts. In this project we will try to monitor osteoblast differentiation under normal and pathological conditions, by means of luciferase-reporter constructs. Retro and/or lenti-viral luciferase-reporter constructs will be made, by cloning the promoter of the osteocalcin, collagen 1A1, and runx2 gene in front of the luciferase gene in a viral vector. Subsequently, freshly isolated MSC or hTert-immortalized MSC cell lines will be transduced with virus-supernatants containing one of these reporter constructs, and will be forced to differentiate into osteoblasts. The amount of luciferase will be used to quantify the differentiation and will be correlated with histological stainings for osteoblast differentiation. Finally, we will try to investigate whether addition of MM cells to the osteoblast cultures results in diminished differentiation and decreased levels of luciferase, and which interaction could be responsible for this phenomenon.
Techniques
Cell culture, FACS analysis/sorting, viral gene marking, molecular imaging (BLI-Fluor) in animal experiments (optional), luciferase assay, liposome production, MSC expansion/differentiation, immuno-histochemistry
Duration
6 or 9 months
Contact
Dr Anton Martens, a.martens@umcutrecht.nl, 088 75 540 09
Dr Kristin Denzer, k.denzer@umcutrecht.nl, 088 75 576 73
More info
UMC Utrecht - dept. of Immunology
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