Application and in vivo tracing of fluorescently labelled stem cells in tissue repair

Far-red fluorescent proteins, e.g. mKate2, E2-Crimson, emit light in the infrared wavelength range. This makes them very useful as markers not only in cultured cells, but also in vivo as they can be detected through non-invasive whole-body imaging of mice (Fluorescence Molecular Tomography (FMT™). Far-red fluorescent proteins can be imaged not only in superficial tissues, but also in organs deep inside the mouse. In a currently on-going joint project in our laboratories, we are labelling mouse embryonic stem (ES-) cells with fluorescent proteins through gene-targeting into the ubiquitously expressed Rosa26 locus, which has been widely used in developmental biology for cell-lineage tracing purposes. After gene targeting, the ES-cells and any cell types derived from them through cell differentiation will express the red fluorescent proteins. The project will employ mKate2 / E2-Crimson labelled stem cells, to investigate stem cell therapies of kidney disease. The far-red labelled ES-cells will be used in vitro, to assess their contribution to tissue-culture models of kidney damage and repair. Furthermore, the fluorescent ES-cells will also be used in vivo in mouse models of kidney regeneration. The project will address the questions of whether and to what extent injected ES-cells contribute to repair and regeneration of damaged kidneys. Additionally, it will investigate ways of partially differentiating fluorescent ES-cells into mesenchymal or kidney precursor cells before application in vivo. The student is invited to join this project, which will comprise a variety of cell culture techniques, molecular biology methods and in vivo analysis of mouse models.