Open AccessDifferentiation and Contractile Analysis of GFP‐Sarcomere Reporter hiPSC‐Cardiomyocytes
Author(s) -
Sharma Arun,
Toepfer Christopher N.,
Schmid Manuel,
Garfinkel Amanda C.,
Seidman Christine E.
Publication year - 2018
Publication title -
current protocols in human genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.282
H-Index - 30
eISSN - 1934-8258
pISSN - 1934-8266
DOI - 10.1002/cphg.53
Subject(s) - sarcomere , induced pluripotent stem cell , contractility , crispr , microbiology and biotechnology , green fluorescent protein , genome editing , myocyte , biology , human induced pluripotent stem cells , embryonic stem cell , gene , genetics , endocrinology
Human induced pluripotent stem cell‐derived cardiomyocytes (hiPSC‐CMs) represent a powerful cellular platform for illuminating mechanisms of human cardiovascular disease and for pharmacological screening. Recent advances in CRISPR/Cas9‐mediated genome editing technology underlie this profound utility. We have generated hiPSC‐CMs harboring fluorescently‐tagged sarcomeric proteins, which provide a tool to non‐invasively study human sarcomere function and dysfunction. In this unit, we illustrate methods for conducting high‐efficiency, small molecule‐mediated differentiation of hiPSCs into cardiomyocytes, and for performing non‐invasive contractile analysis through direct sarcomere tracking of GFP‐sarcomere reporter hiPSC‐CMs. We believe that this type of analysis can overcome sensitivity problems found in other forms of contractile assays involving hiPSC‐CMs by directly measuring contractility at the fundamental contractile unit of the hiPSC‐CM, the sarcomere. © 2018 by John Wiley & Sons, Inc.