An Optimized Protocol for Human Direct Cardiac Reprogramming

Tiffany A Garbutt; Yang Zhou; Benjamin Keepers; Jiandong Liu; Li Qian

doi:10.1016/j.xpro.2019.100010

An Optimized Protocol for Human Direct Cardiac Reprogramming

STAR Protoc. 2020 Jun 19;1(1):100010. doi: 10.1016/j.xpro.2019.100010. Epub 2020 Jun 3.

Authors

Tiffany A Garbutt^{1

2

3

4}, Yang Zhou^{1

2

5

3}, Benjamin Keepers^{1

2}, Jiandong Liu^{1

2}, Li Qian^{1

2

6}

Affiliations

¹ Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, NC 27599, USA.
² McAllister Heart Institute, University of North Carolina, Chapel Hill, NC 27599, USA.
³ These authors contributed equally.
⁴ Technical Contact.
⁵ Current Address: Department of Biomedical Engineering, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
⁶ Lead Contact.

Abstract

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).

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cell Culture Techniques
Cell Separation
Cellular Reprogramming*
Cytological Techniques*
Fibroblasts / cytology*
Humans
MicroRNAs
Myocytes, Cardiac / cytology*
Myocytes, Cardiac / metabolism
Troponin T / biosynthesis

Substances

MicroRNAs
Troponin T

Abstract

Publication types

MeSH terms

Substances

Grants and funding