A pre-requisite for cell differentiation in a 3D environment with subsequent tissue formation is a
coordinated contact formation, together with defined interactions and communication between the
cells. In order to study cellular interactions, one is currently limited to sectioning dead tissue or to ex-
vivo co-culture. The static aspect of non-living cells allows no dynamic view, and in a co-culturing
approach within a provided matrix, cells are embedded randomly. Hence, cell-cell contacts form at
arbitrary areas within the 3D matrix. Therefore, analysis of molecular inter-population interactions is
more difficult. Moreover, culturing of cells restricted to two dimensions is known to alter the cellular
pheno/genotype yielding different behavior, polarity, protein distribution, and cell signaling. Hence, a
system that enables the molecular characterization of cell adhesion, communication and in particular
differentiation, like for e.g. during an osteogenesis, is inevitably needed at the single-cell level in the
context of tissue formation.
The main focus of LiSSCeD will be the development of three-dimensional cell scaffolds mimicking the
extracellular matrix. We plan to combine new 3D lithography methods namely multiphoton- and
Stimulated-Emission-Depletion (STED)-lithography to structure polymers in three dimensions from
hundreds of microns down to several nanometers. Thereby, a strict size control of the polymer
nanostructures, their mechanical properties as well as the selective functionalization with
biomolecules mediating mesenchymal stem cell (MSC) differentiation will be achieved. The scaffolds
will be used as a tool for identification and subsequent optimization of conditions required for MSC
osteogenesis.
The project consortium covers all research fields relevant in this project, namely multiphoton
lithography, single molecule microscopy, localization microscopy as well as cell biophysics (expertise
of the Department of Medical Engineering at the University of Applied Sciences). The Institute of
Applied Physics at the JKU provides its expertise in STED-lithography (as the only institution in
Austria), together with surface- and polymer-chemistry. A close cooperation with the Ludwig
Boltzmann Institute for Experimental and Clinical Traumatology (LBI) and the Department of
Obstetrics at the Medical University of Vienna, which both have a long standing experience in stem
cell research and tissue engineering, will help us in cell cultivation within this project.