Measuring the Electrophysiological Activity of Neuronal Networks Using Micro-Electrode Arrays

Abstract

Source: Frega, M. et al. Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays. J. Vis. Exp. (2017).

The video demonstrates how to use a microelectrode array (MEA) to record the electrophysiological activity of neurons derived from human induced pluripotent stem cells (hiPSCs). Electrophysiological activity from a multiwell MEA containing a neuron-astrocyte co-culture is recorded to detect action potentials from multiple neurons in the network, confirming the successful differentiation of hiPSCs into neurons.

Protocol

1. Differentiation of rtTA/Ngn2-positive hiPSCs to Neurons on 6-well MEAs and Glass Coverslips

NOTE: In this protocol, the details are provided for differentiating reverse tetracycline-controlled transactivator (rtTA)/ Neurogenin-2 (Ngn2)-positive hiPSCs on 6-well MEAs (devices composed of 6 independent wells with 9 recording and 1 reference embedded microelectrodes per well).

1. Prepare the MEAs (day 0 and day 1)
   1. The day before the start of the differentiation, sterilize the MEAs according to the manufacturer's re....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
B-27 supplement	Gibco	0080085SA
0.05 % Trypsin-EDTA	Gibco	25300-054
High-glucose DMEM	Gibco	11965-092
FBS	Sigma-Aldrich	F2442-500ML
Penicillin/Streptomycin	Sigma-Aldrich	P4333
DPBS	Gibco	14190-094
DMEM/F12	Gibco	11320-074
Cell detachment solution	Sigma-Aldrich	A6964
E8 medium	Gibco	A1517001
ROCK inhibitor	Gibco	A2644501	Alternatively, ROCK inhibitors like thiazovivin can be used.
6-well MEAs	Multi Channel Systems	60-6wellMEA200/30iR-Ti-tcr
Poly-L-ornithine	Sigma-Aldrich	P3655-10MG
Laminin	Sigma-Aldrich	L2020-1MG
N-2 supplement	Gibco	17502-048
Non-essential amino acids	Sigma-Aldrich	M7145
NT-3, human recombinant	Promokine	C66425
BDNF, human recombinant	Promokine	C66212
Trypsin-EDTA	Gibco	25300-054
L-alanyl-L-glutamine	Gibco	35050-038
Neurobasal medium	Gibco	21103-049
Cytosine β-D-arabinofuranoside	Sigma-Aldrich	C1768-100MG