Transfection of hPSC-Cardiomyocytes Using Viafect™ Transfection Reagent

Bodbin, Sara E.; Denning, Chris; Mosqueira, Diogo

Abstract

Twenty years since their first derivation, human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) have shown promise in disease modelling research, while their potential for cardiac repair is being investigated. However, low transfection efficiency is a barrier to wider realisation of the potential this model system has to offer. We endeavoured to produce a protocol for improved transfection of hPSC-CMs using the ViafectTM reagent by Promega. Through optimisation of four essential parameters: (i) serum supplementation, (ii) time between replating and transfection, (iii) reagent to DNA ratio and (iv) cell density, we were able to successfully transfect hPSC-CMs to ~95% efficiencies. Transfected hPSC-CMs retained high purity and structural integrity despite a mild reduction in viability, and preserved compatibility with phenotyping assays of hypertrophy. This protocol greatly adds value to the field by overcoming limited transfection efficiencies of hPSC-CMs in a simple and quick approach that ensures sustained expression of transfected genes for at least 14 days, opening new opportunities in mechanistic discovery for cardiac-related diseases

Citation

Bodbin, S. E., Denning, C., & Mosqueira, D. (2020). Transfection of hPSC-Cardiomyocytes Using Viafect™ Transfection Reagent. Methods and Protocols, 3(3), Article 57. https://doi.org/10.3390/mps3030057

• Journal Article Type: Article
• Acceptance Date: Aug 6, 2020
• Online Publication Date: Aug 9, 2020
• Publication Date: Aug 9, 2020
• Deposit Date: Aug 25, 2022
• Publicly Available Date: Aug 25, 2022
• Journal: Methods and Protocols
• Publisher: MDPI
• Peer Reviewed: Peer Reviewed
• Volume: 3
• Issue: 3
• Article Number: 57
• DOI: https://doi.org/10.3390/mps3030057
• Public URL: https://nottingham-repository.worktribe.com/output/4824515
• Publisher URL: https://www.mdpi.com/2409-9279/3/3/57