Defined MicroRNAs Induce Aspects of Maturation in Mouse and Human Embryonic-Stem-Cell-Derived Cardiomyocytes
Desy S. Lee,
Jyh-Hong Chen,
David J. Lundy,
Chung-Hung Liu,
Shiaw-Min Hwang,
Lil Pabon,
Ru-Chi Shieh,
Chien-Chang Chen,
Sheng-Nan Wu,
Yu-Ting Yan,
Sho-Tone Lee,
Po-Min Chiang,
Shu Chien,
Charles E. Murry,
Patrick C.H. Hsieh
Affiliations
Desy S. Lee
Institute of Clinical Medicine, National Cheng Kung University, Tainan 704, Taiwan
Jyh-Hong Chen
Department of Medicine, National Cheng Kung University, Tainan 704, Taiwan
David J. Lundy
Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
Chung-Hung Liu
Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
Shiaw-Min Hwang
Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu 300, Taiwan
Lil Pabon
Department of Pathology, Center for Cardiovascular Biology, Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA
Ru-Chi Shieh
Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
Chien-Chang Chen
Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
Sheng-Nan Wu
Department of Physiology, National Cheng Kung University, Tainan 704, Taiwan
Yu-Ting Yan
Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
Sho-Tone Lee
Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
Po-Min Chiang
Institute of Clinical Medicine, National Cheng Kung University, Tainan 704, Taiwan
Shu Chien
Department of Bioengineering, University of California at San Diego, San Diego, CA 92093, USA
Charles E. Murry
Department of Pathology, Center for Cardiovascular Biology, Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA
Patrick C.H. Hsieh
Institute of Clinical Medicine, National Cheng Kung University, Tainan 704, Taiwan
Pluripotent-cell-derived cardiomyocytes have great potential for use in research and medicine, but limitations in their maturity currently constrain their usefulness. Here, we report a method for improving features of maturation in murine and human embryonic-stem-cell-derived cardiomyocytes (m/hESC-CMs). We found that coculturing m/hESC-CMs with endothelial cells improves their maturity and upregulates several microRNAs. Delivering four of these microRNAs, miR-125b-5p, miR-199a-5p, miR-221, and miR-222 (miR-combo), to m/hESC-CMs resulted in improved sarcomere alignment and calcium handling, a more negative resting membrane potential, and increased expression of cardiomyocyte maturation markers. Although this could not fully phenocopy all adult cardiomyocyte characteristics, these effects persisted for two months following delivery of miR-combo. A luciferase assay demonstrated that all four miRNAs target ErbB4, and siRNA knockdown of ErbB4 partially recapitulated the effects of miR-combo. In summary, a combination of miRNAs induced via endothelial coculture improved ESC-CM maturity, in part through suppression of ErbB4 signaling.
