Lactate and Immunomagnetic-purified iPSC-derived Cardiomyocytes Generate Comparable Engineered Cardiac Tissue Constructs

Rossler, K. J.; de Lange, W.; Mann, M. W.; Aballo, T. J.; Melby, J. A.; Zhang, J.; Kim, G.; Bayne, E. F.; Zhu, Y.; Farrell, E. T.; Kamp, T. J.; Ralphe, J. C.; Ge, Y.

doi:10.1101/2023.05.05.539642

bioRxiv · 10.1101/2023.05.05.539642

Lactate and Immunomagnetic-purified iPSC-derived Cardiomyocytes Generate Comparable Engineered Cardiac Tissue Constructs

Rossler, K. J.·de Lange, W.·Mann, M. W.·Aballo, T. J.·Melby, J. A.·Zhang, J.·Kim, G.·Bayne, E. F.·Zhu, Y.·Farrell, E. T.·Kamp, T. J.·Ralphe, J. C.·Ge, Y.

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Abstract

Three-dimensional engineered cardiac tissue (ECT) using purified human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) has emerged as an appealing model system for the study of human cardiac biology and disease. A recent study reported widely-used metabolic (lactate) purification of monolayer hiPSC-CM cultures results in an ischemic cardiomyopathy-like phenotype compared to magnetic antibody-based cell sorting (MACS) purification, complicating the interpretation of studies using lactate-purified hiPSC-CMs. Herein, our objective was to determine if use of lactate relative to MACs-purified hiPSC-CMs impacts the properties of resulting hiPSC-ECTs. Therefore, hiPSC-CMs were differentiated and purified using either lactate-based media or MACS. After purification, hiPSC-CMs were combined with hiPSC-cardiac fibroblasts to create 3D hiPSC-ECT constructs maintained in culture for four weeks. There were no structural differences observed, and there was no significant difference in sarcomere length between lactate and MACS hiPSC-ECTs. Assessment of isometric twitch force, Ca2+ transients, and {beta}-adrenergic response revealed similar functional performance between purification methods. High-resolution mass spectrometry (MS)-based quantitative proteomics showed no significant difference in any protein pathway expression or myofilament proteoforms. Taken together, this study demonstrates lactate- and MACS-purified hiPSC-CMs generate ECTs with comparable molecular and functional properties, and suggests lactate purification does not result in an irreversible change in hiPSC-CM phenotype. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=177 SRC="FIGDIR/small/539642v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@54b370org.highwire.dtl.DTLVardef@d0bbc8org.highwire.dtl.DTLVardef@1d847e3org.highwire.dtl.DTLVardef@578c1b_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Rossler, K. J., de Lange, W., Mann, M. W., Aballo, T. J., Melby, J. A., Zhang, J., Kim, G., Bayne, E. F., Zhu, Y., Farrell, E. T., Kamp, T. J., Ralphe, J. C., Ge, Y.. 2023-05-06. Lactate and Immunomagnetic-purified iPSC-derived Cardiomyocytes Generate Comparable Engineered Cardiac Tissue Constructs. https://doi.org/10.1101/2023.05.05.539642

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