STAR Protocols (Jun 2020)

An Optimized Protocol for Human Direct Cardiac Reprogramming

  • Tiffany A. Garbutt,
  • Yang Zhou,
  • Benjamin Keepers,
  • Jiandong Liu,
  • Li Qian

Journal volume & issue
Vol. 1, no. 1
p. 100010

Abstract

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Summary: Direct cardiac reprogramming, the conversion of fibroblasts into cardiomyocyte-like cells (iCMs), is an attractive approach to heal the injured heart. Here we present a new approach to human cardiac reprogramming that utilizes a polycistronic three-factor reprogramming cocktail and one microRNA. Our protocol produces cardiac Troponin T positive human iCMs (hiCMs) at an efficiency of 40%–60%, approximately double that of previous protocols, within just 2 weeks. The resulting hiCMs display cardiomyocyte-like sarcomere structure, gene expression, and calcium oscillation.For complete details on the use and execution of this protocol, please refer to Zhou et al. (2019).