Engineering human ventricular heart muscles based on a highly efficient system for purification of human pluripotent stem cell-derived ventricular cardiomyocytes

Bin Li, Hui Yang, Xiaochen Wang, Yongkun Zhan, Wei Sheng, Huanhuan Cai, Haoyang Xin, Qianqian Liang, Ping Zhou, Chao Lu, Ruizhe Qian, Sifeng Chen, Pengyuan Yang, Jianyi Zhang, Weinian Shou, Guoying Huang, Ping Liang, Ning Sun

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background: Most infarctions occur in the left anterior descending coronary artery and cause myocardium damage of the left ventricle. Although current pluripotent stem cells (PSCs) and directed cardiac differentiation techniques are able to generate fetal-like human cardiomyocytes, isolation of pure ventricular cardiomyocytes has been challenging. For repairing ventricular damage, we aimed to establish a highly efficient purification system to obtain homogeneous ventricular cardiomyocytes and prepare engineered human ventricular heart muscles in a dish. Methods: The purification system used TALEN-mediated genomic editing techniques to insert the neomycin or EGFP selection marker directly after the myosin light chain 2 (MYL2) locus in human pluripotent stem cells. Purified early ventricular cardiomyocytes were estimated by immunofluorescence, fluorescence-Activated cell sorting, quantitative PCR, microelectrode array, and patch clamp. In subsequent experiments, the mixture of mature MYL2-positive ventricular cardiomyocytes and mesenchymal cells were cocultured with decellularized natural heart matrix. Histological and electrophysiology analyses of the formed tissues were performed 2 weeks later. Results: Human ventricular cardiomyocytes were efficiently isolated based on the purification system using G418 or flow cytometry selection. When combined with the decellularized natural heart matrix as the scaffold, functional human ventricular heart muscles were prepared in a dish. Conclusions: These engineered human ventricular muscles can be great tools for regenerative therapy of human ventricular damage as well as drug screening and ventricular-specific disease modeling in the future.

Original languageEnglish (US)
Article number651
JournalStem Cell Research and Therapy
Volume8
Issue number1
DOIs
StatePublished - Sep 29 2017

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Pluripotent Stem Cells
Human Engineering
Human engineering
Stem cells
Cardiac Myocytes
Purification
Muscle
Myocardium
Electrophysiology
Neomycin
Flow cytometry
Microelectrodes
Clamping devices
Sorting
Scaffolds
Flow Cytometry
Screening
Fluorescence
Cells
Preclinical Drug Evaluations

Keywords

  • Engineered human heart tissues
  • Engineered human ventricular heart muscles
  • Human pluripotent stem cells
  • Human ventricular cardiomyocytes
  • Myosin light chain 2
  • Myosin light chain 2v

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Molecular Medicine
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • Cell Biology

Cite this

Engineering human ventricular heart muscles based on a highly efficient system for purification of human pluripotent stem cell-derived ventricular cardiomyocytes. / Li, Bin; Yang, Hui; Wang, Xiaochen; Zhan, Yongkun; Sheng, Wei; Cai, Huanhuan; Xin, Haoyang; Liang, Qianqian; Zhou, Ping; Lu, Chao; Qian, Ruizhe; Chen, Sifeng; Yang, Pengyuan; Zhang, Jianyi; Shou, Weinian; Huang, Guoying; Liang, Ping; Sun, Ning.

In: Stem Cell Research and Therapy, Vol. 8, No. 1, 651, 29.09.2017.

Research output: Contribution to journalArticle

Li, B, Yang, H, Wang, X, Zhan, Y, Sheng, W, Cai, H, Xin, H, Liang, Q, Zhou, P, Lu, C, Qian, R, Chen, S, Yang, P, Zhang, J, Shou, W, Huang, G, Liang, P & Sun, N 2017, 'Engineering human ventricular heart muscles based on a highly efficient system for purification of human pluripotent stem cell-derived ventricular cardiomyocytes', Stem Cell Research and Therapy, vol. 8, no. 1, 651. https://doi.org/10.1186/s13287-017-0651-x
Li, Bin ; Yang, Hui ; Wang, Xiaochen ; Zhan, Yongkun ; Sheng, Wei ; Cai, Huanhuan ; Xin, Haoyang ; Liang, Qianqian ; Zhou, Ping ; Lu, Chao ; Qian, Ruizhe ; Chen, Sifeng ; Yang, Pengyuan ; Zhang, Jianyi ; Shou, Weinian ; Huang, Guoying ; Liang, Ping ; Sun, Ning. / Engineering human ventricular heart muscles based on a highly efficient system for purification of human pluripotent stem cell-derived ventricular cardiomyocytes. In: Stem Cell Research and Therapy. 2017 ; Vol. 8, No. 1.
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AU - Yang, Hui

AU - Wang, Xiaochen

AU - Zhan, Yongkun

AU - Sheng, Wei

AU - Cai, Huanhuan

AU - Xin, Haoyang

AU - Liang, Qianqian

AU - Zhou, Ping

AU - Lu, Chao

AU - Qian, Ruizhe

AU - Chen, Sifeng

AU - Yang, Pengyuan

AU - Zhang, Jianyi

AU - Shou, Weinian

AU - Huang, Guoying

AU - Liang, Ping

AU - Sun, Ning

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N2 - Background: Most infarctions occur in the left anterior descending coronary artery and cause myocardium damage of the left ventricle. Although current pluripotent stem cells (PSCs) and directed cardiac differentiation techniques are able to generate fetal-like human cardiomyocytes, isolation of pure ventricular cardiomyocytes has been challenging. For repairing ventricular damage, we aimed to establish a highly efficient purification system to obtain homogeneous ventricular cardiomyocytes and prepare engineered human ventricular heart muscles in a dish. Methods: The purification system used TALEN-mediated genomic editing techniques to insert the neomycin or EGFP selection marker directly after the myosin light chain 2 (MYL2) locus in human pluripotent stem cells. Purified early ventricular cardiomyocytes were estimated by immunofluorescence, fluorescence-Activated cell sorting, quantitative PCR, microelectrode array, and patch clamp. In subsequent experiments, the mixture of mature MYL2-positive ventricular cardiomyocytes and mesenchymal cells were cocultured with decellularized natural heart matrix. Histological and electrophysiology analyses of the formed tissues were performed 2 weeks later. Results: Human ventricular cardiomyocytes were efficiently isolated based on the purification system using G418 or flow cytometry selection. When combined with the decellularized natural heart matrix as the scaffold, functional human ventricular heart muscles were prepared in a dish. Conclusions: These engineered human ventricular muscles can be great tools for regenerative therapy of human ventricular damage as well as drug screening and ventricular-specific disease modeling in the future.

AB - Background: Most infarctions occur in the left anterior descending coronary artery and cause myocardium damage of the left ventricle. Although current pluripotent stem cells (PSCs) and directed cardiac differentiation techniques are able to generate fetal-like human cardiomyocytes, isolation of pure ventricular cardiomyocytes has been challenging. For repairing ventricular damage, we aimed to establish a highly efficient purification system to obtain homogeneous ventricular cardiomyocytes and prepare engineered human ventricular heart muscles in a dish. Methods: The purification system used TALEN-mediated genomic editing techniques to insert the neomycin or EGFP selection marker directly after the myosin light chain 2 (MYL2) locus in human pluripotent stem cells. Purified early ventricular cardiomyocytes were estimated by immunofluorescence, fluorescence-Activated cell sorting, quantitative PCR, microelectrode array, and patch clamp. In subsequent experiments, the mixture of mature MYL2-positive ventricular cardiomyocytes and mesenchymal cells were cocultured with decellularized natural heart matrix. Histological and electrophysiology analyses of the formed tissues were performed 2 weeks later. Results: Human ventricular cardiomyocytes were efficiently isolated based on the purification system using G418 or flow cytometry selection. When combined with the decellularized natural heart matrix as the scaffold, functional human ventricular heart muscles were prepared in a dish. Conclusions: These engineered human ventricular muscles can be great tools for regenerative therapy of human ventricular damage as well as drug screening and ventricular-specific disease modeling in the future.

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KW - Myosin light chain 2v

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