Protocol to measure calcium spikes in cardiomyocytes obtained from human pluripotent stem cells using a ready-to-use media

Veronica Astro; Gustavo Ramirez-Calderon; Antonio Adamo

doi:10.1016/j.xpro.2023.102252

Protocol to measure calcium spikes in cardiomyocytes obtained from human pluripotent stem cells using a ready-to-use media

STAR Protoc. 2023 Apr 13;4(2):102252. doi: 10.1016/j.xpro.2023.102252. Online ahead of print.

Authors

Veronica Astro¹, Gustavo Ramirez-Calderon², Antonio Adamo³

Affiliations

¹ Biological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia. Electronic address: veronica.astro@kaust.edu.sa.
² Biological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia.
³ Biological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia. Electronic address: antonio.adamo@kaust.edu.sa.

Abstract

The derivation of cardiomyocytes from human pluripotent stem cells (hPSCs) is a powerful tool to investigate early cardiogenesis and model diseases in vitro. Here, we present an optimized protocol to obtain contracting hPSCs-derived cardiomyocytes using a ready-to-use kit. We describe steps for hPSC culture and differentiation to cardiomyocytes including the identification of key parameters such as starting cell confluency and temperature. We then detail immunofluorescence, flow cytometry, and the quantification of cardiomyocytes' calcium spikes using live imaging. For complete details on the use and execution of this protocol, please refer to Astro et al.¹.

Keywords: Cell Biology; Cell Culture; Cell Differentiation; Flow Cytometry/Mass Cytometry; Microscopy; Stem Cells.