Mesenchymal stem cells in live cell chips for high-content screenings in functional genomics

^1,2Christian Maercker, ³Tilmann Rogge, ⁴Heidrun Ridinger, ⁵Karen Bieback
¹German Cancer Researach Center (DKFZ), D-69120 Heidelberg, Germany, ²Mannheim University of Applied Sciences, D-68163 Mannheim, Germany, ³Helmholtz Research Center Karlsruhe, D-76344 Eggenstein-Leopoldshafen, Germany, ⁴RZPD German Resource Center for Genome Research, D-69120 Heidelberg, Germany, ⁵University Hospital Mannheim, D-68167 Mannheim, Germany

There is a big demand for high-throughput functional assays which can measure molecular phenotypes and therefore contribute to networks in systems biology, making them applicable in the investigation of complex diseases and drug target validation. Our high-content screening approach focuses on adhesion as an important differentiation process which is monitored to investigate effects of pathogenic factors or drug leads, respectively. Different cell types were cultivated in plastic chips with integrated electrodes to measure impedance or capacitance as a rate of cell adhesion. In parallel, protein biomarkers have been followed by fluorescence microscopy. It has been possible to discriminate between different cell densities and cell types. Moreover, after stimulation human cardiac fibroblasts and human umbilical vein endothelial cells, concentrations of proteins involved in adhesion have been increased after stimulation and proteins were translocated within the cells. In ongoing experiments, fibroblastoid mesenchymal stem cells are isolated from fat tissue, umbilical cord, or bone marrow. To optimize adhesion conditions, the surfaces within the vials of the chip are specifically activated. Microscopy is adjusted to be able to measure cellular morphology in parallel. Our concept allows to identify mesenchymal stem cell's behavior, which can not be described by standard biomarkers so far. In addition, simulation of the homing process of the cells within its stem cell niche in an in vitro assay is a promising setup for large-scale gain-of-function or loss-of-function screenings in functional genomics.