Micropatterning of substrates for the culture of cell networks by stencil-assisted additive nanofabrication

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Previdi, Anita and Piazzoni, Claudio and Borghi, Francesca and Schulte, Carsten and Lorenzelli, Leandro and Giacomozzi, Flavio and Bucciarelli, Alessio and Malgaroli, Antonio and Lamanna, Jacopo and Moro, Andrea and Racchetti, Gabriella and Podestà, Alessandro and Lenardi, Cristina and Milani, Paolo (2021) Micropatterning of substrates for the culture of cell networks by stencil-assisted additive nanofabrication. Micromachines, 12 (1). 94. ISSN 2072-666X (https://doi.org/10.3390/mi12010094)

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Abstract

The fabrication of in vitro neuronal cell networks where cells are chemically or electrically connected to form functional circuits with useful properties is of great interest. Standard cell culture substrates provide ensembles of cells that scarcely reproduce physiological structures since their spatial organization and connectivity cannot be controlled. Supersonic Cluster Beam Deposition (SCBD) has been used as an effective additive method for the large-scale fabrication of interfaces with extracellular matrix-mimicking surface nanotopography and reproducible morphological properties for cell culture. Due to the high collimation of SCBD, it is possible to exploit stencil masks for the fabrication of patterned films and reproduce features as small as tens of micrometers. Here, we present a protocol to fabricate micropatterned cell culture substrates based on the deposition of nanostructured cluster-assembled zirconia films by stencil-assisted SCBD. The effectiveness of this approach is demonstrated by the fabrication of micrometric patterns able to confine primary astrocytes. Calcium waves propagating in the astrocyte networks are shown.

ORCID iDs

Previdi, Anita, Piazzoni, Claudio, Borghi, Francesca, Schulte, Carsten ORCID logoORCID: https://orcid.org/0000-0002-7554-5342, Lorenzelli, Leandro, Giacomozzi, Flavio, Bucciarelli, Alessio, Malgaroli, Antonio, Lamanna, Jacopo, Moro, Andrea, Racchetti, Gabriella, Podestà, Alessandro, Lenardi, Cristina and Milani, Paolo;
CORE (COnnecting REpositories)