A Novel 3D Label-Free Monitoring System of hES-Derived Cardiomyocyte Clusters: A Step Forward to In Vitro Cardiotoxicity Testing
Publication Date
July 08, 2013
Journal
PLoS ONE
Authors
Heinz-Georg Jahnke, Daniella Steel, Stephan Fleischer, Diana Seidel, et al
Volume
8
Issue
7
Pages
e68971
DOI
https://dx.plos.org/10.1371/journal.pone.0068971
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0068971
Web of Science
000321692000034
Scopus
84879968282
Mendeley
http://www.mendeley.com/research/novel-3d-labelfree-monitoring-system-hesderived-cardiomyocyte-clusters-step-forward-vitro-cardiotoxi
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Mendeley | Further Information

{"title"=>"A Novel 3D Label-Free Monitoring System of hES-Derived Cardiomyocyte Clusters: A Step Forward to In Vitro Cardiotoxicity Testing", "type"=>"journal", "authors"=>[{"first_name"=>"Heinz Georg", "last_name"=>"Jahnke", "scopus_author_id"=>"8532600400"}, {"first_name"=>"Daniella", "last_name"=>"Steel", "scopus_author_id"=>"36656336400"}, {"first_name"=>"Stephan", "last_name"=>"Fleischer", "scopus_author_id"=>"55788067000"}, {"first_name"=>"Diana", "last_name"=>"Seidel", "scopus_author_id"=>"55470377800"}, {"first_name"=>"Randy", "last_name"=>"Kurz", "scopus_author_id"=>"8532600100"}, {"first_name"=>"Silvia", "last_name"=>"Vinz", "scopus_author_id"=>"55789215600"}, {"first_name"=>"Kerstin", "last_name"=>"Dahlenborg", "scopus_author_id"=>"6603311232"}, {"first_name"=>"Peter", "last_name"=>"Sartipy", "scopus_author_id"=>"6701750291"}, {"first_name"=>"Andrea A.", "last_name"=>"Robitzki", "scopus_author_id"=>"6603901754"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203", "scopus"=>"2-s2.0-84879968282", "sgr"=>"84879968282", "pui"=>"369286874", "doi"=>"10.1371/journal.pone.0068971", "pmid"=>"23861955", "issn"=>"19326203"}, "id"=>"d7108b18-db6a-3499-9128-c055538ad97e", "abstract"=>"Unexpected adverse effects on the cardiovascular system remain a major challenge in the development of novel active pharmaceutical ingredients (API). To overcome the current limitations of animal-based in vitro and in vivo test systems, stem cell derived human cardiomyocyte clusters (hCMC) offer the opportunity for highly predictable pre-clinical testing. The three-dimensional structure of hCMC appears more representative of tissue milieu than traditional monolayer cell culture. However, there is a lack of long-term, real time monitoring systems for tissue-like cardiac material. To address this issue, we have developed a microcavity array (MCA)-based label-free monitoring system that eliminates the need for critical hCMC adhesion and outgrowth steps. In contrast, feasible field potential derived action potential recording is possible immediately after positioning within the microcavity. Moreover, this approach allows extended observation of adverse effects on hCMC. For the first time, we describe herein the monitoring of hCMC over 35 days while preserving the hCMC structure and electrophysiological characteristics. Furthermore, we demonstrated the sensitive detection and quantification of adverse API effects using E4031, doxorubicin, and noradrenaline directly on unaltered 3D cultures. The MCA system provides multi-parameter analysis capabilities incorporating field potential recording, impedance spectroscopy, and optical read-outs on individual clusters giving a comprehensive insight into induced cellular alterations within a complex cardiac culture over days or even weeks.", "link"=>"http://www.mendeley.com/research/novel-3d-labelfree-monitoring-system-hesderived-cardiomyocyte-clusters-step-forward-vitro-cardiotoxi", "reader_count"=>34, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>14, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>1, "Student > Bachelor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>14, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>1, "Student > Bachelor"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>8, "Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>13, "Medicine and Dentistry"=>5, "Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>8}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>13}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Switzerland"=>2, "Germany"=>1}, "group_count"=>2}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1113630"], "description"=>"<p>(A) The recorded field potential of one hCMC at the beginning (control) and after application of 100 µM noradrenaline. The comparison of the concentration response curve for the accumulative (n  = 6 hCMC) and discrete application (n  = 3 hCMC) of noradrenaline reveals considerably different EC<sub>50</sub> values. (B) Analysis of adverse side effects like QT interval relevant APD prolongation can be detected and quantified. While E4031 has an influence on contraction rate at higher concentrations, the application of 1 nM results in a prolonged fAPD after 15 minutes. The analysis of the frequency-corrected fAPD (fAPD<sub>c</sub>) revealed an EC<sub>50</sub> of 7.2 nM (n  = 13 hCMC), (blue bar – published EC<sub>50</sub> range obtained from hERG-channel assay, green bar – published EC<sub>10</sub> range of <i>in vivo</i> studies in dogs, monkeys and humans).</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Embryonic stem cells", "Molecular cell biology", "Cellular types", "toxicology", "Predictive toxicology", "cardiovascular", "Drugs and devices", "Cardiovascular pharmacology", "Drug research and development", "Real-time", "detection", "api", "adverse"], "article_id"=>741384, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Heinz-Georg Jahnke", "Daniella Steel", "Stephan Fleischer", "Diana Seidel", "Randy Kurz", "Silvia Vinz", "Kerstin Dahlenborg", "Peter Sartipy", "Andrea A. Robitzki"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068971.g003", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_hCMC_for_real_time_detection_of_API_adverse_effects_/741384", "title"=>"hCMC for real-time detection of API adverse effects.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-08 01:35:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/1113628"], "description"=>"<p>Comparative analysis of available techniques (monolayer cultures and hCMC attached to planar MEA) and the MCA technology. (A) Analysis of contractility and contraction rate stability for each method over 35 days. (B) Analysis of fAPD occurrence and fAPD stability for each method over 35 days. (C) Representative examples of recorded field potentials from the three methods.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Embryonic stem cells", "Molecular cell biology", "Cellular types", "toxicology", "Predictive toxicology", "cardiovascular", "Drugs and devices", "Cardiovascular pharmacology", "Drug research and development", "improves", "potential-derived"], "article_id"=>741382, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Heinz-Georg Jahnke", "Daniella Steel", "Stephan Fleischer", "Diana Seidel", "Randy Kurz", "Silvia Vinz", "Kerstin Dahlenborg", "Peter Sartipy", "Andrea A. Robitzki"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068971.g002", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MCA_technology_improves_quality_and_long_term_stability_of_field_potential_derived_action_potential_parameters_/741382", "title"=>"MCA technology improves quality and long-term stability of field potential-derived action potential parameters.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-08 01:35:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/1113636"], "description"=>"<p>(A) The contraction rate was analyzed for 48 hours (n  = 4 hCMC). The effect on the APD was determined for one hour after doxorubicin application and revealed a concentration dependent response with an EC<sub>50</sub> value of 17 µM (n  = 3 hCMC), (B) The impedance analysis revealed a concentration- and time-dependent decrease where IC<sub>50</sub> values could be determined for each time point (n  = 5 hCMC). (C) The microscopic analysis of the hCMC size (cross section area) showed an increase only for 100 µM after 24 hours and 48 hours (n  = 5). (D) After the experiment, the clusters were suitable for further molecular biological analysis. The immunocytochemical staining indicated typical stratification of contractile elements (arrows) in the control and 0.1 µM treated sample. These stratified elements were not observed in samples treated with 1 µM doxorubicin (bar  = 50 µm). For comparison with established <i>in vivo</i> cardiomyocyte damage markers, a c-troponin assay was performed on the culture media.</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Embryonic stem cells", "Molecular cell biology", "Cellular types", "toxicology", "Predictive toxicology", "cardiovascular", "Drugs and devices", "Cardiovascular pharmacology", "Drug research and development", "Real-time", "multi-parametric", "monitoring", "enables"], "article_id"=>741390, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Heinz-Georg Jahnke", "Daniella Steel", "Stephan Fleischer", "Diana Seidel", "Randy Kurz", "Silvia Vinz", "Kerstin Dahlenborg", "Peter Sartipy", "Andrea A. Robitzki"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068971.g004", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Label_free_real_time_multi_parametric_monitoring_enables_high_content_analysis_/741390", "title"=>"Label-free real-time multi-parametric monitoring enables high content analysis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-08 01:35:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/1113623"], "description"=>"<p>(A) Bonded silicone-based microcavity array and SEM image of a cavity (400 µm size) with an integrated suction hole for automated positioning. (B) Microcavity array technology enables an instant and detailed field potential recording on four electrodes per cluster without any adhesion requirement of the cardiomyocyte cluster to the electrodes. (C) The hCMC consist of highly enriched cardiomyocytes with atrial (MLC-2a) and to a smaller amount of ventricular cardiomyocyte characteristics (MLC-2v), (bar  = 100 µm, zoom: bar  = 20 µm). (D) Size and electrophysiological characteristics of six independent differentiation experiments each with 36 hCMC (mean ± s.d.).</p>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Embryonic stem cells", "Molecular cell biology", "Cellular types", "toxicology", "Predictive toxicology", "cardiovascular", "Drugs and devices", "Cardiovascular pharmacology", "Drug research and development", "microcavity", "purified", "cardiomyocyte"], "article_id"=>741377, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Heinz-Georg Jahnke", "Daniella Steel", "Stephan Fleischer", "Diana Seidel", "Randy Kurz", "Silvia Vinz", "Kerstin Dahlenborg", "Peter Sartipy", "Andrea A. Robitzki"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068971.g001", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Optimized_microcavity_array_for_field_potential_analysis_of_highly_purified_cardiomyocyte_clusters_/741377", "title"=>"Optimized microcavity array for field potential analysis of highly purified cardiomyocyte clusters.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-07-08 01:35:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/1113639", "https://ndownloader.figshare.com/files/1113641", "https://ndownloader.figshare.com/files/1113642", "https://ndownloader.figshare.com/files/1113644", "https://ndownloader.figshare.com/files/1113645", "https://ndownloader.figshare.com/files/1113646", "https://ndownloader.figshare.com/files/1113647", "https://ndownloader.figshare.com/files/1113648", "https://ndownloader.figshare.com/files/1113650", "https://ndownloader.figshare.com/files/1113651"], "description"=>"<div><p>Unexpected adverse effects on the cardiovascular system remain a major challenge in the development of novel active pharmaceutical ingredients (API). To overcome the current limitations of animal-based <i>in vitro</i> and in vivo test systems, stem cell derived human cardiomyocyte clusters (hCMC) offer the opportunity for highly predictable pre-clinical testing. The three-dimensional structure of hCMC appears more representative of tissue milieu than traditional monolayer cell culture. However, there is a lack of long-term, real time monitoring systems for tissue-like cardiac material. To address this issue, we have developed a microcavity array (MCA)-based label-free monitoring system that eliminates the need for critical hCMC adhesion and outgrowth steps. In contrast, feasible field potential derived action potential recording is possible immediately after positioning within the microcavity. Moreover, this approach allows extended observation of adverse effects on hCMC. For the first time, we describe herein the monitoring of hCMC over 35 days while preserving the hCMC structure and electrophysiological characteristics. Furthermore, we demonstrated the sensitive detection and quantification of adverse API effects using E4031, doxorubicin, and noradrenaline directly on unaltered 3D cultures. The MCA system provides multi-parameter analysis capabilities incorporating field potential recording, impedance spectroscopy, and optical read-outs on individual clusters giving a comprehensive insight into induced cellular alterations within a complex cardiac culture over days or even weeks.</p></div>", "links"=>[], "tags"=>["developmental biology", "stem cells", "Embryonic stem cells", "Molecular cell biology", "Cellular types", "toxicology", "Predictive toxicology", "cardiovascular", "Drugs and devices", "Cardiovascular pharmacology", "Drug research and development", "3d", "label-free", "monitoring", "hes-derived", "cardiomyocyte", "cardiotoxicity"], "article_id"=>741393, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Heinz-Georg Jahnke", "Daniella Steel", "Stephan Fleischer", "Diana Seidel", "Randy Kurz", "Silvia Vinz", "Kerstin Dahlenborg", "Peter Sartipy", "Andrea A. Robitzki"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0068971.s001", "https://dx.doi.org/10.1371/journal.pone.0068971.s002", "https://dx.doi.org/10.1371/journal.pone.0068971.s003", "https://dx.doi.org/10.1371/journal.pone.0068971.s004", "https://dx.doi.org/10.1371/journal.pone.0068971.s005", "https://dx.doi.org/10.1371/journal.pone.0068971.s006", "https://dx.doi.org/10.1371/journal.pone.0068971.s007", "https://dx.doi.org/10.1371/journal.pone.0068971.s008", "https://dx.doi.org/10.1371/journal.pone.0068971.s009", "https://dx.doi.org/10.1371/journal.pone.0068971.s010"], "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/A_Novel_3D_Label_Free_Monitoring_System_of_hES_Derived_Cardiomyocyte_Clusters_A_Step_Forward_to_In_Vitro_Cardiotoxicity_Testing/741393", "title"=>"A Novel 3D Label-Free Monitoring System of hES-Derived Cardiomyocyte Clusters: A Step Forward to <i>In Vitro</i> Cardiotoxicity Testing", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-07-08 01:35:00"}

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Relative Metric

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