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Efficient and reproducible generation of human iPSC-derived cardiomyocytes using a stirred bioreactor. | LitMetric

AI Article Synopsis

  • Human iPSC-derived cardiomyocytes (hiPSC-CMs) are important for studying cardiac diseases and regeneration, but issues with scaling and consistency hinder their use in research and clinical settings.
  • A new cardiac differentiation protocol using Wnt modulation and a stirred suspension bioreactor successfully produces high-purity hiPSC-CMs, averaging 124 million cells per batch with excellent viability and consistent properties across different cell lines.
  • Compared to traditional monolayer-derived cardiomyocytes (mCMs), these bioreactor-derived cardiomyocytes (bCMs) exhibit better structural and functional characteristics, including enhanced sarcomere maturation, stronger force production in 3D heart tissues, and improved viability, making them more suitable for various applications

Article Abstract

In the last decade human iPSC-derived cardiomyocytes (hiPSC-CMs) proved to be valuable for cardiac disease modeling and cardiac regeneration, yet challenges with scale, quality, inter-batch consistency, and cryopreservation remain, reducing experimental reproducibility and limiting clinical translation. Here, we report a robust cardiac differentiation protocol that uses Wnt modulation and a stirred suspension bioreactor to produce on average 124 million hiPSC-CMs with >90% purity using a variety of hiPSC lines (19 differentiations; 10 iPSC lines). After controlled freeze and thaw, bioreactor-derived CMs (bCMs) showed high viability (>90%), interbatch reproducibility in cellular morphology, function, drug response and ventricular identity, which was further supported by single cell transcriptomes. bCMs on microcontact printed substrates revealed a higher degree of sarcomere maturation and viability during long-term culture compared to monolayer-derived CMs (mCMs). Moreover, functional investigation of bCMs in 3D engineered heart tissues showed earlier and stronger force production during long-term culture, and robust pacing capture up to 4 Hz when compared to mCMs. bCMs derived from this differentiation protocol will expand the applications of hiPSC-CMs by providing a reproducible, scalable, and resource efficient method to generate cardiac cells with well-characterized structural and functional properties superior to standard mCMs.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10925150PMC
http://dx.doi.org/10.1101/2024.02.24.581789DOI Listing

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