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An Optimized Protocol for Human Direct Cardiac Reprogramming
Tiffany A Garbutt1,2,3,4, Yang Zhou1,2,5,3, Benjamin Keepers1,2
1Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, NC 27599, USA.
STAR Protocols
|July 31, 2020
Summary
This study introduces an efficient new method for direct cardiac reprogramming, converting fibroblasts into cardiomyocyte-like cells (hiCMs) in just two weeks. The novel protocol achieves high efficiency, offering a promising approach for heart repair strategies.
Area of Science:
- Cardiovascular Biology
- Stem Cell Biology
- Regenerative Medicine
Background:
- Direct cardiac reprogramming offers a potential therapeutic strategy for cardiac repair by converting somatic cells into cardiomyocyte-like cells.
- Existing protocols for human cardiac reprogramming face limitations in efficiency and time.
Purpose of the Study:
- To develop a novel and highly efficient protocol for direct cardiac reprogramming of human fibroblasts into cardiomyocyte-like cells (hiCMs).
- To characterize the efficiency and functional properties of hiCMs generated by the new protocol.
Main Methods:
- Utilized a polycistronic three-factor reprogramming cocktail combined with a specific microRNA.
- Applied the protocol to human fibroblasts for direct conversion into hiCMs.
Main Results:
- Achieved a 40%-60% efficiency in generating cardiac Troponin T-positive hiCMs within 2 weeks.
- The generated hiCMs exhibited cardiomyocyte-like sarcomere structure, gene expression profiles, and functional calcium oscillations.
- The efficiency was approximately double that of previously reported protocols.
Conclusions:
- The novel reprogramming protocol significantly enhances the efficiency and speed of generating functional hiCMs.
- This optimized direct cardiac reprogramming method holds promise for future applications in regenerative cardiology and treating heart disease.
