Toward physiological conditions for cell analyses: forces of heart muscle cells suspended between elastic micropillars

Almost each mammalian cell permanently applies forces to its environment. These forces are essential for many vital processes such as tissue formation or cell movement. In turn, the environmental conditions of cells strongly affect force production. Here we report on the development of an array of elastomeric micropillars as cellular environment. Within these micropillar arrays, we cultivated rat heart muscle cells (cardiac myocytes). For lattice constants between 20 and 30 μm, cells strongly preferred spanning between the elastic micropillars over adhering to the underlying flat substrate. In addition, the architectures of the cytoskeleton and of protein complexes formed for adhesion were strongly dependent on the environment of the cell. On flat parts of the substrates, we observed prominent stress fibers and focal adhesion sites. In contrast, cells suspended between micropillars exhibited well organized myofibers and costameric adhesions at the locations of Z-bands. These observations argue for close-to-nature environmental conditions within micropillar arrays. Resting as well as contraction forces of myocytes resulted in measurable pillar bending. Using an approximate theoretical treatment of elastically founded micropillars, we calculated average cell forces of 140 nN in the relaxed and 400 nN in the contracted state.