A chip-based platform for the in vitro generation of tissues in three-dimensional organization

Abstract

We describe a multi-purpose platform for the three-dimensional cultivation of tissues. The device is composed of polymer chips featuring a microstructured area of 1-2 cm(2). The chip is constructed either as a grid of micro-containers measuring 120-300 x 300 x 300 microm (h x l x w), or as an array of round recesses (300 microm diameter, 300 microm deep). The micro-containers may be separately equipped with addressable 3D-micro-electrodes, which allow for electrical stimulation of excitable cells and on-site measurements of electrochemically accessible parameters. The system is applicable for the cultivation of high cell densities of up to 8 x 10(6) cells and, because of the rectangular grid layout, allows the automated microscopical analysis of cultivated cells. More than 1000 micro-containers enable the parallel analysis of different parameters under superfusion/perfusion conditions. Using different polymer chips in combination with various types of bioreactors we demonstrated the principal suitability of the chip-based bioreactor for tissue culture applications. Primary and established cell lines have been successfully cultivated and analysed for functional properties. When cells were cultured in non-perfused chips, over time a considerable degree of apoptosis could be observed indicating the need for an active perfusion. The system presented here has also been applied for the differentiation analysis of pluripotent embryonic stem cells and may be suitable for the analysis of the stem cell niche.

Chip-based Three-dimensional Cell Culture in Perfused Micro-bioreactors

“…Depending on the experimental design, the surface of the polymer can be modified by UV-irradiation 4 . Hepatocyte cell lines as well as primary rat hepatocytes can successfully be cultivated in these devices as could be shown by expression analysis of some liver specific genes as well as analysis of some liver specific proteins 5,6 .…”

Microstructuring of multiwell plates for three-dimensional cell culture applications by ultrasonic embossing

“…This chip has a central grid-like microstructured area with cubic microcavities in which cells can organize into uniform multicellular aggregates (Gottwald et al 2007;Weibezahn et al 1993). The cavities of the chip are open to the top and have a side length of 300 μm in each direction (w × l × d).…”

“…In a second step, a solvent-vapor-welding technique is used to bond a porous membrane to the open bottom of the chip. As the geometry of the chip array proved to be suitable for the generation of uniform adherent multicellular aggregates (Altmann et al 2008;Gottwald et al 2007;Rieke et al 2008), our intention was to transfer the chip design to multiwell cell culture plates for HTS applications.…”

The famousversusthe inconvenient - or the dawn and the rise of 3D-culture systems

“…Finally, electrospinning offers new 3D scaffolds with double length scale features with combinations of microfibers and electrospun nanofibers [37] . In contrast to the above mentioned techniques, we use a culture system developed at the Karlsruhe Institute of Technology that is based on a microstructured polymer chip that serves as a scaffold for the 3D cultivation of cells [38][39][40][41] . Currently, the chip is manufactured in two different designs varying mainly in geometry and the manufacturing process.…”