Switchable cell adhesive microstructures to grow defined neuronal networks
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1
University of Rostock, Department of Biophysics, Germany
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2
Fraunhofer-Institut für Zelltherapie und Immunologie (IZI), Branch Bioanalytics and Bioprocesses, Germany
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3
Berlin-Brandenburgisches Zentrum für Regenerative Therapien, Charité Universitätsmedizin, Germany
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4
Freie Universität Berlin, Germany
The molecular structure of TRPs (thermo-responsive polymers) alters below and above a certain temperature. In experiments with neuronal cell lines (SH-SY5Y), we used TRP, which repelled cells below 33 °C and favored cell adhesion above 37 °C. For a more detailed characterization of the TRP properties, we determined the initial cell adhesion forces with single cell force spectroscopy. For the temporal and spatial control of the formation of neuronal circuits in vitro, micro-patterned structures with thermo-responsive surfaces were developed.
Glass chips were produced with heating micro-structures covered by a Si3N4 passivation layer and coated with gold (Figure 1, gold). Experiments with primary neuronal mouse cells, which did not favor direct adhesion to TRP surfaces required micro-structuration of the TRP surfaces. Thiol-gold chemistry was used to coat the chip surfaces with TRP. The gold surface featured micro-structured round spots with protruding trenches, which prevented TRP adsorption but permitted the adhesion of single cells directly at the Si3N4 passivation layer (Figure 1, blue). The trenches, which connected the cell adhesion spots served to guide the axons of the cells to their neighbors (Figure 1, green). Cell adhesion to the feeder areas, trenches and cell adhesion spots could be enhanced by their specific surface modification with (3-aminopropyl)-triethoxysilane. The cell spots were connected to feeder areas on both sides of the chips (Figure 1, mint green). A gap in the trench to each neighboring cell prevented the direct connection of the axons to the neighbor cells (Figure 1, red). Heating micro-structures were located underneath the gaps. They permitting to switch between the cell adhesive and non-adhesive states of the TRP cover layer. Our final goal is the cultivation of primary neurons in defined network structures and the control of the connection in cellular arrays.
Figure 1: Micro-structured chip for an array of four cells with two heating structures, four cell adhesion spot, and guiding trenches (blue: single cell adhesion spot, green: axon guide, red: thermo-switchable gap, gold: TRP-coated gold, mint green: feeder area). Scale bar: 100 µm.
Acknowledgements
The authors are grateful to the DFG (German Research Council) for funding the Projekt "NeuroTRP" (#290023374)
Keywords:
Thermo-responsive polymers,
neuronal circuits,
primary neuronal cells,
Directed growth,
microsystems
Conference:
MEA Meeting 2018 | 11th International Meeting on Substrate Integrated Microelectrode Arrays, Reutlingen, Germany, 4 Jul - 6 Jul, 2018.
Presentation Type:
Poster Presentation
Topic:
Neural Networks
Citation:
Wysotzki
P,
Schröder
J,
Behm
LV,
Gerike
S,
Pfisterer
F,
Duschl
C,
Kirschbaum
M,
Gimsa
J and
Baumann
W
(2019). Switchable cell adhesive microstructures to grow defined neuronal networks.
Conference Abstract:
MEA Meeting 2018 | 11th International Meeting on Substrate Integrated Microelectrode Arrays.
doi: 10.3389/conf.fncel.2018.38.00102
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Received:
15 Mar 2018;
Published Online:
17 Jan 2019.
*
Correspondence:
Mr. Philipp Wysotzki, University of Rostock, Department of Biophysics, Rostock, Germany, 18057, Germany, Philipp.wysotzki@uni-rostock.de