Pluripotent Stem-Cell Derived Cardiomyocytes

This volume provides methodologies for ES and iPS cell technology on the study of cardiovascular diseases. Chapters guide readers through protocols on cardiomyocyte generation from pluripotent stem cells, physiological measurements, bioinformatic analysis, gene editing technology, and cell transplantation studies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

 

Authoritative and cutting-edge, Pluripotent Stem-Cell Derived Cardiomyocytes aims to help researchers set up experiments using pluripotent stem cell-derived cardiac cells.

Part I: Overview

1. Making Cardiomyocytes from Pluripotent Stem Cells      

Peter Karaginius and Yoshinori Yoshida

 

Part II: Generation of Pluripotent Stem Cell-Derived Cardiomyocytes and Cardiac Tissues

2. A method for Large-Scale Cardiac Differentiation, Purification, and Cardiac Spheroid Production of Human Induced Pluripotent Stem Cells         

Yuika Morita, Shugo Tohyama, Jun Fujita, and Keiichi Fukuda

 

3. Large-scale Differentiation of Stem Cell-Derived Cardiomyocytes by Stirring-Type Suspension Culture          

Nagako Sougawa, Shigeru Miyagawa, and Yoshiki Sawa

 

4. Efficient Method to Dissociate Induced Pluripotent Stem Cell-derived Cardiomyocyte Aggregates into Single Cells   

Emiko Ito, Shigeru Miyagawa, Yoshinori Yoshida, and Yoshiki Sawa

 

5. Isolation of Cardiomyocytes Derived from Human Pluripotent Stem Cells using miRNA switches       

Kenji Miki, Hirohide Saito, and Yoshinori Yoshida

 

6. Fabrication of Cardiac Constructs using bio-3D Printer   

Kenichi Arai, Daiki Murata, Shoko Takao, and Koichi Nakayama

 

7. Fabrication of Thick and Anisotropic Cardiac Tissue on Nanofibrous Substrate for Repairing Infarcted Myocardium   

Junjun Li, Li Liu, Itsunari Minami, Shigeru Miyagawa, and Yoshiki Sawa

 

8. Construction of Three-Dimensional Cardiac Tissues Using Layer by Layer Method       

Maki Takeda, Shigeru Miyagawa, Mitsuru Akashi, and Yoshiki Sawa

 

9. Generation of Cylindrical Engineered Cardiac Tissues from human iPS Cell-derived Cardiovascular Cell Lineages           

Hidetoshi Masumoto

 

Part III: Physiological Mesurements using Pluripotent Stem Cell-Derived Cardiomyocytes

 

10. Protocol for Morphological and Functional Phenotype Analysis of hiPS-derived Cardiomyocytes      

Jun LI and Jong-Kook LEE

 

11. Application of FluoVolt Membrane Potential Dye for Induced Pluripotent Stem Cell-derived Cardiac Single Cells and Monolayers Differentiated via Embryoid Bodies 

Tadashi Takaki and Yoshinori Yoshida

 

12. Multi-Electrode Array Assays Using Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes           

Daisuke Yoshinaga, Yimin Wuriyanghai, and Takeru Makiyama

 

13. Electrophysiological Analysis of hiPSC-derived Cardiomyocytes Using a Patch-Clamp Technique     

Yuta Yamamoto, Sayako Hirose, Yimin Wuriyanghai, Daisuke Yoshinaga, and Takeru Makiyama

 

14. Characterization of Ventricular and Atrial Cardiomyocyte Subtypes from Human Induced Pluripotent Stem Cells           

Misato Koakutsu, Tadashi Takaki, Kenji Miki, and Yoshinori Yoshida

 

15. Assessment of Contractility in Human iPS Cell-Derived Cardiomyocytes Using Motion Vector Analysis       

Yasunari Kanda, Ayano Satsuka, Sayo Hayashi, Mihoko Hagiwara-Nagasawa, and Atsushi Sugiyama

 

16. Contractile Force Measurement of Engineered Cardiac Tissues Derived from Human iPS Cells

Daisuke Sasaki, Katsuhisa Matsuura, and Tatsuya Shimizu

 

17. A Method for Contraction Force Measurement of iPSC Derived Engineered Cardiac Tissues

Yuya Fujiwara, Kohei Deguchi, Kenji Miki, Tomoyuki Nishimoto, and Yoshinori Yoshida

 

Part IV: Transcriptome and Bioinfomatics Analysis

18. Single-cardiomyocyte RNA Sequencing to Dissect the Molecular Pathophysiology of the Heart

Manami Kotoh, Seitaro Nomura, Shintaro Yamada,Hiroyuki Aburatani, and Issei Komuro

 

19. RNA-sequencing Analysis of Differentially Expressed Genes in Human iPSC-derived Cardiomyocytes        

Chikako Okubo, Megumi Narita, Takuya Yamamoto, and Yoshinori Yoshida

 

20. Analysis of Transcriptional Profiling of Chamber-specific Human Cardiac Myocytes Derived from Pluripotent Stem Cells

Antonio Lucena-Cacace and Yoshinori Yoshida

 

Part V: Gene Editing and CRISPR Technology for Pluripotent Stem Cells

21. Genome Editing in Human Induced Pluripotent Stem Cells (hiPSCs)   

Shuichiro Higo, Shungo Hikoso, Shigeru Miyagawa, and Yasushi Sakata

 

22. Generation of Efficient Knock-in Mouse and Human Pluripotent Stem Cells Using CRISPR-Cas9    

Tatsuya Anzai, Hiromasa Hara, Chanthra Nawin, Taketaro Sadahiro, Masaki Ieda, Yutaka Hanazono, and Hideki Uosaki

 

23. CRISPRi/a Screening with Human iPSCs           

Masataka Nishiga, Lei S. Qi, and Joseph C. Wu

 

Part VI: Transplantation of Pluripotent Stem Cell-Derived Cardiomyocytes

24. Transplantation of Human Induced Pluripotent Stem Cell-derived Cardiomyocytes in a Mouse Myocardial Infarction Model        

Takeshi Hatani and Yoshinori Yoshida

 

25. Transplantation of Pluripotent Stem Cell-derived Cardiomyocytes into a Myocardial Infarction Model of Cynomolgus Monkey 

Hideki Kobayashi, Hajime Ichimura, Noburou Ohashi, and Yuji Shiba