Accurate nanoelectrode recording of human pluripotent stem cell-derived cardiomyocytes for assaying drugs and modeling disease

Ziliang Carter Lin,Hsin-Ya Lou,Paul W Burridge,Allister F Mcguire,Joseph C Wu,Bianxiao Cui,Elena Matsa

doi:10.1038/micronano.2016.80

Abstract

The measurement of the electrophysiology of human pluripotent stem cell-derived cardiomyocytes is critical for their biomedical applications, from disease modeling to drug screening. Yet, a method that enables the high-throughput intracellular electrophysiology measurement of single cardiomyocytes in adherent culture is not available. To address this area, we have fabricated vertical nanopillar electrodes that can record intracellular action potentials from up to 60 single beating cardiomyocytes. Intracellular access is achieved by highly localized electroporation, which allows for low impedance electrical access to the intracellular voltage. Herein, we demonstrate that this method provides the accurate measurement of the shape and duration of intracellular action potentials, validated by patch clamp, and can facilitate cellular drug screening and disease modeling using human pluripotent stem cells. This study validates the use of nanopillar electrodes for myriad further applications of human pluripotent stem cell-derived cardiomyocytes such as cardiomyocyte maturation monitoring and electrophysiology-contractile force correlation.

Highlights

From repairing damaged tissue in vivo to predicting drug efficacy, human pluripotent stem cells have become a promising solution to many bottlenecks in human biological research[1,2,3,4,5,6]
We observed a dramatic amplitude increase and a signal shape that matches that of patch clamp-recorded intracellular action potentials
The second disease we studied was long QT syndrome (LQTS), which is characterized by the delayed repolarization and prolongation of the action potential and may be studied using human induced pluripotent stem cells (hiPSCs)-CMs28

Summary

Introduction

From repairing damaged tissue in vivo to predicting drug efficacy, human pluripotent stem cells (hPSCs) have become a promising solution to many bottlenecks in human biological research[1,2,3,4,5,6]. There have been many efforts to generate cardiomyocytes (CMs) derived from human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) due to the difficulty in obtaining human cardiac tissue by other methods[7]. HPSC-CMs exhibit key cardiomyocyte properties, expressing the expected ion channels, exhibiting human cardiac-type action potentials, and containing functional sarcomeres. Despite their potential, applications of hPSC-CMs have been hampered by the cells’ intrinsic heterogeneity and the quality of functional assays for monitoring their electrophysiology. The high heterogeneity among hPSC-CMs requires that a large number of cells be analyzed to generate statistically meaningful conclusions

Methods

Results

Conclusion