Efficient and reproducible generation of human iPSC-derived cardiomyocytes and cardiac organoids in stirred suspension systems

Maksymilian Prondzynski, Paul Berkson, Michael A. Trembley, Yashasvi Tharani, Kevin Shani, Raul H. Bortolin, Mason E. Sweat, Joshua Mayourian, Dogacan Yucel, Albert M. Cordoves, Beatrice Gabbin, Cuilan Hou, Nnaemeka J. Anyanwu, Farina Nawar, Justin Cotton, Joseph Milosh, David Walker, Yan Zhang, Fujian Lu, Xujie Liu, Kevin Kit Parker, Vassilios J. Bezzerides and William T. Pu ()
Additional contact information
Maksymilian Prondzynski: Boston Children’s Hospital
Paul Berkson: Boston Children’s Hospital
Michael A. Trembley: Boston Children’s Hospital
Yashasvi Tharani: Boston Children’s Hospital
Kevin Shani: Harvard University
Raul H. Bortolin: Boston Children’s Hospital
Mason E. Sweat: Boston Children’s Hospital
Joshua Mayourian: Boston Children’s Hospital
Dogacan Yucel: Boston Children’s Hospital
Albert M. Cordoves: Harvard University
Beatrice Gabbin: Boston Children’s Hospital
Cuilan Hou: Boston Children’s Hospital
Nnaemeka J. Anyanwu: Harvard University
Farina Nawar: Boston Children’s Hospital
Justin Cotton: Boston Children’s Hospital
Joseph Milosh: Boston Children’s Hospital
David Walker: Boston Children’s Hospital
Yan Zhang: Boston Children’s Hospital
Fujian Lu: Boston Children’s Hospital
Xujie Liu: Boston Children’s Hospital
Kevin Kit Parker: Harvard University
Vassilios J. Bezzerides: Boston Children’s Hospital
William T. Pu: Boston Children’s Hospital

Nature Communications, 2024, vol. 15, issue 1, 1-17

Abstract: Abstract Human iPSC-derived cardiomyocytes (hiPSC-CMs) have proven invaluable for cardiac disease modeling and regeneration. Challenges with quality, inter-batch consistency, cryopreservation and scale remain, reducing experimental reproducibility and clinical translation. Here, we report a robust stirred suspension cardiac differentiation protocol, and we perform extensive morphological and functional characterization of the resulting bioreactor-differentiated iPSC-CMs (bCMs). Across multiple different iPSC lines, the protocol produces 1.2E6/mL bCMs with ~94% purity. bCMs have high viability after cryo-recovery (>90%) and predominantly ventricular identity. Compared to standard monolayer-differentiated CMs, bCMs are more reproducible across batches and have more mature functional properties. The protocol also works with magnetically stirred spinner flasks, which are more economical and scalable than bioreactors. Minor protocol modifications generate cardiac organoids fully in suspension culture. These reproducible, scalable, and resource-efficient approaches to generate iPSC-CMs and organoids will expand their applications, and our benchmark data will enable comparison to cells produced by other cardiac differentiation protocols.

Date: 2024
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DOI: 10.1038/s41467-024-50224-0

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