Cells and Viruses
The Cells and Viruses platform uses various cell systems to specifically implement and analyze genetic changes in cells using viral systems. Among other things, this is done using recombinant adeno-associated viruses (AAVs) and retroviruses. In addition to in vitro applications, these are also used for in vivo applications. This can be used to modify the expression of a specific gene, for example Egfl7 (overexpression or knockout). By means of AAV-induced Cre recombinase expression, this can also be a tissue-specific (for example, hippocampus) as well as promoter-driven cell type-specific knockout. In addition, applications of AAVs and retroviruses in vivo can be used to specifically track cell populations or perform axon tracing.
Furthermore, adherent cells, neurospheres, co-cultures and suspension cultures are used for further downstream analyses. Cell-cell interaction studies, cell adhesion experiments, analyses of migration and invasion capacity by means of Boyden chambers or proliferation and apoptosis studies are applied. In addition to basic cell culture techniques, complex cultivation systems such as slice cultures of murine brains and brain regions (e.g. hippocampus) are used.
In addition, our platform is able to culture and analyze primary neurospheres. For this purpose, subpopulations of the neural lineage are also sorted in a highly purified manner using fluorescence-activated cell sorting (FACS). This allows, for example, the separate cultivation of neural stem cells and proliferating progenitor cells, so that the influence on individual cell stages can be investigated. These sorted cells are also available for further downstream analysis, such as RNA sequencing.
Furthermore, cells of different culture conditions or primary tissue are analyzed for the expression of specific surface molecules by flow cytometry on the BD FACSymphony™ A3 Cell Analyzer (FACS facility, MTZ). A variety of panels using up to 23 markers for the different cell types (such as immune cells and their subpopulations, or glioma cells) are employed.