At the present state of the art, CSCs are isolated from a tumor sample, such as cancer cell lines, via flow cytometry to obtain sufficiently pure cancer stem cells for consecutive experiments. After purification, asymmetric division causes that the percentage of CSCs declines with each cell division to finally result in a frequency of 1% or less. This commonly limits the timeframe and the scale of consecutive experiments. Accordingly, there are intense international efforts to generate cancer cells, which are "arrested" in the CSC-phenotype and that can be propagated infinitely or at least over longer time spans with already some initial success reported in the recent literature.
The main goal of this subproject is to systematically explore genes and miRNAs known or suspected to be related to the CSC-phenotype for their ability to "arrest" cancer cells in the CSC-state and to consecutively characterize these cells. Furthermore, a broader panel of cell lines is generated, in which the CSCs are marked with different reporter constructs in order to trace their individual fates in a complex population.