Enhancing human pluripotent stem cell differentiation to cardiomyocytes through cardiac progenitor reseeding and cryopreservation
Year: 2025
Type: article
Abstract: Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) have the potential to transform the understanding of heart development and heart failure treatment. However, hPSC-CM differentiation efficiency is plagued by batch-to-batch and line-to-line variability. Here, we describe a method to improve CM purity by 10–20% (absolute) without negatively affecting contractility, sarcomere structure, multinucleation, junctional Cx43, or CM number by detaching and reseeding progenitors between the EOMES + mesoderm and ISL1 +/ NKX2-5 + cardiac progenitor stages. Moreover, we demonstrate that EOMES + mesoderm and ISL1 +/ NKX2-5 + cardiac progenitors are cryopreservable with similar improvements in CM purity after resuming differentiation, facilitating storage of large batches of hPSC-CM progenitors for on-demand CM production. Reseeding during differentiation also enables transition to defined extracellular matrices, including fibronectin, vitronectin, and laminin-111, which all supported hPSC-derived EOMES + mesoderm and ISL1 +/ NKX2-5 + cardiac progenitor differentiation to CMs. In summary, we present a method to increase hPSC-CM differentiation purity and demonstrate that specific CM progenitors are amenable to cryopreservation.
Source: iScience
Institution: University of Wisconsin–Madison
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,240
Awards: G2375471656, G406957865, G4849603943, NSF Engineering Research Center for Cell Manufacturing Technologies (CMaT), G6786228269, Enhancing robustness of human pluripotent stem cell differentiation to cardiomyocytes by uncovering on- and off-target differentiation trajectories via single nucleus multi-omics, G7036742205, G7127918317, Biotechnology Training Program
OpenAlex ID: W4409548416