Previdi Anita, Piazzoni Claudio, Borghi Francesca, Schulte Carsten, Lorenzelli Leandro, Giacomozzi Flavio, Bucciarelli Alessio, Malgaroli Antonio, Lamanna Jacopo, Moro Andrea, Racchetti Gabriella, Podestà Alessandro, Lenardi Cristina, Milani Paolo
CIMaINa and Dipartimento di Fisica, Università degli Studi di Milano, Via Celoria 16, 20133 Milano, Italy.
Center for Materials and Microsystems (CMM), Bruno Kessler Foundation (FBK), Via Sommarive 18, 38123 Trento, Italy.
Micromachines (Basel). 2021 Jan 18;12(1):94. doi: 10.3390/mi12010094.
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.
构建体外神经元细胞网络很受关注，在该网络中细胞通过化学或电连接形成具有有用特性的功能电路。标准细胞培养基质提供的细胞集合几乎无法重现生理结构，因为其空间组织和连接性无法控制。超声团簇束沉积（SCBD）已被用作一种有效的添加方法，用于大规模制造具有模仿细胞外基质表面纳米拓扑结构和可重现形态特性的界面以进行细胞培养。由于SCBD的高准直性，利用模板掩膜制造图案化薄膜并重现小至几十微米的特征成为可能。在此，我们展示了一种基于模板辅助SCBD沉积纳米结构团簇组装氧化锆薄膜来制造微图案化细胞培养基质的方案。通过制造能够限制原代星形胶质细胞的微米级图案证明了该方法的有效性。展示了在星形胶质细胞网络中传播的钙波。
