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Human iPSC-based cardiac microphysiological system for drug screening applications.

Mathur A, Loskill P, Shao K, Huebsch N, Hong S, Marcus SG, Marks N, Mandegar M, Conklin BR, Lee LP, Healy KE - Sci Rep (2015)

Bottom Line: Drug discovery and development are hampered by high failure rates attributed to the reliance on non-human animal models employed during safety and efficacy testing.Pharmacological studies using the cardiac MPS show half maximal inhibitory/effective concentration values (IC₅₀/EC₅₀) that are more consistent with the data on tissue scale references compared to cellular scale studies.We anticipate the widespread adoption of MPSs for drug screening and disease modeling.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Bioengineering and California Institute for Quantitative Biosciences (QB3), University of California at Berkeley, Berkeley, California 94720, USA [2] Department of Materials Science and Engineering, University of California at Berkeley, Berkeley, California 94720, USA.

ABSTRACT
Drug discovery and development are hampered by high failure rates attributed to the reliance on non-human animal models employed during safety and efficacy testing. A fundamental problem in this inefficient process is that non-human animal models cannot adequately represent human biology. Thus, there is an urgent need for high-content in vitro systems that can better predict drug-induced toxicity. Systems that predict cardiotoxicity are of uppermost significance, as approximately one third of safety-based pharmaceutical withdrawals are due to cardiotoxicty. Here, we present a cardiac microphysiological system (MPS) with the attributes required for an ideal in vitro system to predict cardiotoxicity: i) cells with a human genetic background; ii) physiologically relevant tissue structure (e.g. aligned cells); iii) computationally predictable perfusion mimicking human vasculature; and, iv) multiple modes of analysis (e.g. biological, electrophysiological, and physiological). Our MPS is able to keep human induced pluripotent stem cell derived cardiac tissue viable and functional over multiple weeks. Pharmacological studies using the cardiac MPS show half maximal inhibitory/effective concentration values (IC₅₀/EC₅₀) that are more consistent with the data on tissue scale references compared to cellular scale studies. We anticipate the widespread adoption of MPSs for drug screening and disease modeling.

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Characterization of the 3D cardiac tissue in the microphysiological system (MPS).(a) Optical microscopy image of a cardiac tissue in the MPS. (b) Confocal fluorescence microscopy imaging of the microtissue in the MPS reveals an aligned multiple cell layer thick structure. Inset shows a magnified view of the sarcomeric alpha actinin and DAPI staining. (c, d) Heat map of the time-averaged beating motion and corresponding average beating kinetics in the MPS, (e, f) along the channel axis, and (g, h) perpendicular to the channel axis.
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f2: Characterization of the 3D cardiac tissue in the microphysiological system (MPS).(a) Optical microscopy image of a cardiac tissue in the MPS. (b) Confocal fluorescence microscopy imaging of the microtissue in the MPS reveals an aligned multiple cell layer thick structure. Inset shows a magnified view of the sarcomeric alpha actinin and DAPI staining. (c, d) Heat map of the time-averaged beating motion and corresponding average beating kinetics in the MPS, (e, f) along the channel axis, and (g, h) perpendicular to the channel axis.

Mentions: The hiPSC-CMs formed a 3D cardiac tissue in serum-free media within 24 hours of loading, which started to beat spontaneously with robust and homogenous beating at physiological beat rates of 55–80 bpm without any external stimulation (Fig. 2a, Supplementary Movie 3). Within 7 days of formation, the beating became aligned in an uniaxial manner and consisted of multiple cell layers, as confirmed by confocal fluorescence microscopy (Fig. 2b). Computational motion-tracking of the beating velocity indicated the beating was aligned along the long axis of the MPS (Fig. 2 c–h). Aligned 3D cell structure and consistent physiological beating are crucial prerequisites for a cardiac MPS, since functionality is dictated by structural alignment of the cells. Drug response, for instance, is strongly dependent on the initial spontaneous beat rate of the tissue21 and the electromechanical activity of the heart is governed by the intracellular connections and 3D alignment of the cardiac tissue2223. The MPS system is highly scalable and presents the opportunity to model human genetic disease via the use of patient-derived or genome-edited hiPSC-CMs in a rapid and cost effective manner. For example, we used hiPSC-CMs derived from a genetically engineered hiPSC line expressing a GCaMP6 reporter to monitor Ca2+ currents21, which provides the ability to collect electrophysiological data on a high-content basis using simple optical microscopy, as shown in Fig. 3 and Supplementary Movie 4.


Human iPSC-based cardiac microphysiological system for drug screening applications.

Mathur A, Loskill P, Shao K, Huebsch N, Hong S, Marcus SG, Marks N, Mandegar M, Conklin BR, Lee LP, Healy KE - Sci Rep (2015)

Characterization of the 3D cardiac tissue in the microphysiological system (MPS).(a) Optical microscopy image of a cardiac tissue in the MPS. (b) Confocal fluorescence microscopy imaging of the microtissue in the MPS reveals an aligned multiple cell layer thick structure. Inset shows a magnified view of the sarcomeric alpha actinin and DAPI staining. (c, d) Heat map of the time-averaged beating motion and corresponding average beating kinetics in the MPS, (e, f) along the channel axis, and (g, h) perpendicular to the channel axis.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4352848&req=5

f2: Characterization of the 3D cardiac tissue in the microphysiological system (MPS).(a) Optical microscopy image of a cardiac tissue in the MPS. (b) Confocal fluorescence microscopy imaging of the microtissue in the MPS reveals an aligned multiple cell layer thick structure. Inset shows a magnified view of the sarcomeric alpha actinin and DAPI staining. (c, d) Heat map of the time-averaged beating motion and corresponding average beating kinetics in the MPS, (e, f) along the channel axis, and (g, h) perpendicular to the channel axis.
Mentions: The hiPSC-CMs formed a 3D cardiac tissue in serum-free media within 24 hours of loading, which started to beat spontaneously with robust and homogenous beating at physiological beat rates of 55–80 bpm without any external stimulation (Fig. 2a, Supplementary Movie 3). Within 7 days of formation, the beating became aligned in an uniaxial manner and consisted of multiple cell layers, as confirmed by confocal fluorescence microscopy (Fig. 2b). Computational motion-tracking of the beating velocity indicated the beating was aligned along the long axis of the MPS (Fig. 2 c–h). Aligned 3D cell structure and consistent physiological beating are crucial prerequisites for a cardiac MPS, since functionality is dictated by structural alignment of the cells. Drug response, for instance, is strongly dependent on the initial spontaneous beat rate of the tissue21 and the electromechanical activity of the heart is governed by the intracellular connections and 3D alignment of the cardiac tissue2223. The MPS system is highly scalable and presents the opportunity to model human genetic disease via the use of patient-derived or genome-edited hiPSC-CMs in a rapid and cost effective manner. For example, we used hiPSC-CMs derived from a genetically engineered hiPSC line expressing a GCaMP6 reporter to monitor Ca2+ currents21, which provides the ability to collect electrophysiological data on a high-content basis using simple optical microscopy, as shown in Fig. 3 and Supplementary Movie 4.

Bottom Line: Drug discovery and development are hampered by high failure rates attributed to the reliance on non-human animal models employed during safety and efficacy testing.Pharmacological studies using the cardiac MPS show half maximal inhibitory/effective concentration values (IC₅₀/EC₅₀) that are more consistent with the data on tissue scale references compared to cellular scale studies.We anticipate the widespread adoption of MPSs for drug screening and disease modeling.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Bioengineering and California Institute for Quantitative Biosciences (QB3), University of California at Berkeley, Berkeley, California 94720, USA [2] Department of Materials Science and Engineering, University of California at Berkeley, Berkeley, California 94720, USA.

ABSTRACT
Drug discovery and development are hampered by high failure rates attributed to the reliance on non-human animal models employed during safety and efficacy testing. A fundamental problem in this inefficient process is that non-human animal models cannot adequately represent human biology. Thus, there is an urgent need for high-content in vitro systems that can better predict drug-induced toxicity. Systems that predict cardiotoxicity are of uppermost significance, as approximately one third of safety-based pharmaceutical withdrawals are due to cardiotoxicty. Here, we present a cardiac microphysiological system (MPS) with the attributes required for an ideal in vitro system to predict cardiotoxicity: i) cells with a human genetic background; ii) physiologically relevant tissue structure (e.g. aligned cells); iii) computationally predictable perfusion mimicking human vasculature; and, iv) multiple modes of analysis (e.g. biological, electrophysiological, and physiological). Our MPS is able to keep human induced pluripotent stem cell derived cardiac tissue viable and functional over multiple weeks. Pharmacological studies using the cardiac MPS show half maximal inhibitory/effective concentration values (IC₅₀/EC₅₀) that are more consistent with the data on tissue scale references compared to cellular scale studies. We anticipate the widespread adoption of MPSs for drug screening and disease modeling.

Show MeSH
Related in: MedlinePlus