Building Three-Dimensional Neuronal Networks Coupled to Micro-Electrode Arrays
Published: July 31, 2024
Abstract
Source: Tedesco, M., et al., Interfacing 3D Engineered Neuronal Cultures to Micro-Electrode Arrays: An Innovative In Vitro Experimental Model. J. Vis. Exp. (2015)
This video demonstrates the development of a 3D neuronal culture coupled to a planar microelectrode array with hippocampal neurons. The isolated neurons form a 2D network on the microelectrode array's active area and adhere to glass microbeads in a multi-well plate with a membrane insert. Upon transferring these neuron-coated microbeads to the electrode, they organize into an interconnected 3D neural network.
Protocol
All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. 1. Dissection of Embryos and Dissociation of Tissue House adult female rats (200-250 g) at a constant temperature (22 ± 1 °C) and relative humidity (50%) under a regular light–dark schedule (light-on 7 AM–7 PM) in the animal facility. Ensure that food and water are freely available. Anesthetize a pregnant female rat a…
Divulgaciones
The authors have nothing to disclose.
Materials
| Laminin | Sigma-Aldrich | L2020 | 0.05 µg/ml |
| Poly-D-lysine | Sigma-Aldrich | P6407 | 0.05 µg/ml |
| Trypsin | Gibco | 25050-014 | 0.125% diluted 1:2 in HBSS wo calcium and magnesium ions |
| DNAase | Sigma-Aldrich | D5025 | 0.05% diluted in Hanks solution |
| Neurobasal | Gibco Invitrogen | 21103049 | Culture medium |
| B27 | Gibco Invitrogen | 17504044 | 2% medium supplent |
| Fetal bovine serum (FBS) | Sigma-Aldrich | F-2442 | 10% |
| Glutamax-I | Gibco | 35050038 | 0.5 mM |
| gentamicin | Sigma-Aldrich | G.1272 | 5mg/liter |
| Poly-Dimethyl-Siloxane (PDMS) | Corning Sigma | 481939 | Curing agent and the polymer |
| Micro-Electrode Arrays | Multi Channel Systems (MCS) | 60MEA200/30-Ti-pr | MEA with: Electrode grid 8×8; Electrode spacing and diameter 200 and 30 µm, respectively; plastic ring without thread |
| Microbead | Distrilab-Duke Scientific | 9040 | 1gr Glass Part.Size Stds 40 µm |
| Transwell | Costar Sigma | CLS 3413 | Multiwell plates with membrane insert (6.5 mm diameter porous 0.4 µm) |
| HBSS wo calcium and magnesium ions | Gibco Invitrogen | 14175-052 | |
| Hanks Buffer Solution | Sigma | H8264 | |
| Rat | Sprague Dawley | Wistar Rat |
Citar este artículo
Building Three-Dimensional Neuronal Networks Coupled to Micro-Electrode Arrays. J. Vis. Exp. (Pending Publication), e22354, doi: (2024).