Ryota Tabei1, Shinji Kawaguchi2, Hideaki Kanazawa3, Shugo Tohyama4, Akinori Hirano2, Noriko Handa1, Shuji Hishikawa5, Takumi Teratani5, Satoshi Kunita5, Junichi Fukuda6, Yoshihiro Mugishima6, Tsuneyoshi Suzuki6, Kazuaki Nakajima1, Tomohisa Seki1, Yoshikazu Kishino1, Marina Okada1, Masataka Yamazaki2, Kazuma Okamoto2, Hideyuki Shimizu2, Eiji Kobayashi7, Yasuhiko Tabata8, Jun Fujita9, Keiichi Fukuda1. 1. Department of Cardiology, Keio University School of Medicine, Tokyo, Japan. 2. Department of Cardiovascular Surgery, Keio University School of Medicine, Tokyo, Japan. 3. Department of Cardiology, Keio University School of Medicine, Tokyo, Japan. Electronic address: kanazawa@a5.keio.jp. 4. Department of Cardiology, Keio University School of Medicine, Tokyo, Japan; Department of Organ Fabrication, Keio University School of Medicine, Tokyo, Japan. 5. Center for Development of Advanced Medical Technology, Jichi Medical University, Tochigi, Japan. 6. Suzuki Precision Co., Ltd., Tochigi, Japan. 7. Department of Organ Fabrication, Keio University School of Medicine, Tokyo, Japan. 8. Department of Biomaterials, Field of Tissue Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan. 9. Department of Cardiology, Keio University School of Medicine, Tokyo, Japan. Electronic address: jfujita@a6.keio.jp.
Abstract
BACKGROUND: Induced pluripotent stem cell (iPSC)‒based regenerative therapy is a promising strategy for cardiovascular disease treatment; however, the method is limited by the myocardial retention of grafted iPSCs. Thus, an injection protocol that efficiently introduces and retains human iPSC-derived cardiomyocytes (hiPSC-CMs) within the myocardium is urgently needed. The objective of the present study was to develop a method to improve the retention of hiPSCs in the myocardium for cardiac therapy. METHODS: We efficiently produced hiPSC-CM spheroids in 3-dimensional (3D) culture using microwell plates, and developed an injection device for optimal 3D distribution of the spheroids in the myocardial layer. Device biocompatibility was assessed with purified hiPSC-CM spheroids. Device effectiveness was evaluated in 10- to 15-month-old farm pigs (n = 15) and 5- to 24-month-old micro-minipigs (n = 20). The pigs were euthanized after injection, and tissues were harvested for retention and histologic analysis. RESULTS: We demonstrated an injection device for direct intramyocardial transplantation of hiPSC-CM spheroids from large-scale culture. The device had no detrimental effects on cell viability, spheroid shape, or size. Direct epicardial injection of spheroids mixed with gelatin hydrogel into beating porcine hearts using this device resulted in better distribution and retention of transplanted spheroids in a layer within the myocardium than did conventional needle injection procedures. CONCLUSIONS: The combination of the newly developed transplant device and spheroid formation promotes the retention of transplanted CMs. These findings support the clinical application of hiPSC-CM spheroid‒based cardiac regenerative therapy in patients with heart failure.
BACKGROUND: Induced pluripotent stem cell (iPSC)‒based regenerative therapy is a promising strategy for cardiovascular disease treatment; however, the method is limited by the myocardial retention of grafted iPSCs. Thus, an injection protocol that efficiently introduces and retains human iPSC-derived cardiomyocytes (hiPSC-CMs) within the myocardium is urgently needed. The objective of the present study was to develop a method to improve the retention of hiPSCs in the myocardium for cardiac therapy. METHODS: We efficiently produced hiPSC-CM spheroids in 3-dimensional (3D) culture using microwell plates, and developed an injection device for optimal 3D distribution of the spheroids in the myocardial layer. Device biocompatibility was assessed with purified hiPSC-CM spheroids. Device effectiveness was evaluated in 10- to 15-month-old farm pigs (n = 15) and 5- to 24-month-old micro-minipigs (n = 20). The pigs were euthanized after injection, and tissues were harvested for retention and histologic analysis. RESULTS: We demonstrated an injection device for direct intramyocardial transplantation of hiPSC-CM spheroids from large-scale culture. The device had no detrimental effects on cell viability, spheroid shape, or size. Direct epicardial injection of spheroids mixed with gelatin hydrogel into beating porcine hearts using this device resulted in better distribution and retention of transplanted spheroids in a layer within the myocardium than did conventional needle injection procedures. CONCLUSIONS: The combination of the newly developed transplant device and spheroid formation promotes the retention of transplanted CMs. These findings support the clinical application of hiPSC-CM spheroid‒based cardiac regenerative therapy in patients with heart failure.
Authors: Marbod Weber; Andreas Fech; Luise Jäger; Heidrun Steinle; Louisa Bühler; Regine Mariette Perl; Petros Martirosian; Roman Mehling; Dominik Sonanini; Wilhelm K Aicher; Konstantin Nikolaou; Christian Schlensak; Markus D Enderle; Hans Peter Wendel; Walter Linzenbold; Meltem Avci-Adali Journal: Sci Rep Date: 2020-10-08 Impact factor: 4.379
Authors: Pallavi Pushp; Diogo E S Nogueira; Carlos A V Rodrigues; Frederico C Ferreira; Joaquim M S Cabral; Mukesh Kumar Gupta Journal: Stem Cell Rev Rep Date: 2020-10-23 Impact factor: 5.739