Home › Cell Biology › Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays
Cell Biology JoVE (Open Access) Citable · DOI

Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays

DOI: 10.3791/54900-v
What you'll learn
  • ✓ Differentiate hiPSCs into excitatory cortical neurons reproducibly
  • ✓ Control neuronal cell density for experimental precision
  • ✓ Measure network-level electrophysiology using micro-electrode arrays
  • ✓ Characterize neuronal activity and network formation kinetics
Protocol

We modify and implement a previously published protocol describing the rapid, reproducible, and efficient differentiation of human induced Pluripotent Stem Cells (hiPSCs) into excitatory cortical neurons12. Specifically, our modification allows for control of neuronal cell density and use on micro-electrode arrays to measure electrophysiological properties at the network level.

Difficulty
advanced
Total time
~14–21 days (hiPSC expansion and differentiation protocol)
Model organism
Human induced pluripotent stem cells (hiPSCs); rat primary astrocytes
Biosafety
BSL-1

Steps

1
Plate hiPSCs on differentiation substrate

Seed hiPSCs at controlled density on prepared culture plates to initiate neural differentiation. Cell density at plating determines final neuronal yield and network properties.

▶ 00:41
2
Add rat astrocytes to support neuronal maturation

Co-culture rat primary astrocytes with differentiating hiPSCs to enhance neuronal maturation and network formation. Astrocytes provide trophic support and improve physiological properties.

▶ 03:42
3
Measure network activity on micro-electrode arrays

Record spontaneous electrophysiological activity from the neuronal network using MEA technology. Quantify firing rates, bursting patterns, and network synchronization in real time.

▶ 05:48
4
Analyze and interpret hiPSC-derived neuronal phenotype

Characterize key features of differentiated neurons including morphology, connectivity, and electrophysiological properties using MEA readouts. Validate protocol reproducibility and cell identity.

▶ 06:31
💬 Comments coming soon