Identification and Characterization of Calcium Sparks in Cardiomyocytes Derived from Human Induced Pluripotent Stem Cells
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{"title"=>"Identification and Characterization of Calcium Sparks in Cardiomyocytes Derived from Human Induced Pluripotent Stem Cells", "type"=>"journal", "authors"=>[{"first_name"=>"Guang Qin", "last_name"=>"Zhang", "scopus_author_id"=>"7405272049"}, {"first_name"=>"Heming", "last_name"=>"Wei", "scopus_author_id"=>"7402516954"}, {"first_name"=>"Jun", "last_name"=>"Lu", "scopus_author_id"=>"57191504071"}, {"first_name"=>"Philip", "last_name"=>"Wong", "scopus_author_id"=>"55231463900"}, {"first_name"=>"Winston", "last_name"=>"Shim", "scopus_author_id"=>"8773017000"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"23408964", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0055266", "pui"=>"368311254", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "scopus"=>"2-s2.0-84873577242", "sgr"=>"84873577242"}, "id"=>"a546b543-7d2e-33e6-9cb8-494ca9c09433", "abstract"=>"INTRODUCTION Ca2+ spark constitutes the elementary units of cardiac excitation-contraction (E-C) coupling in mature cardiomyocytes. Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes are known to have electrophysiological properties similar to mature adult cardiomyocytes. However, it is unclear if they share similar calcium handling property. We hypothesized that Ca2+ sparks in human induced pluripotent stem cell (hiPSCs)-derived cardiomyocytes (hiPSC-CMs) may display unique structural and functional properties than mature adult cardiomyocytes. METHODS AND RESULTS Ca2+ sparks in hiPSC-CMs were recorded with Ca2+ imaging assay with confocal laser scanning microscopy. Those sparks were stochastic with a tendency of repetitive occurrence at the same site. Nevertheless, the spatial-temporal properties of Ca2+ spark were analogous to that of adult CMs. Inhibition of L-type Ca2+ channels by nifedipine caused a 61% reduction in calcium spark frequency without affecting amplitude of those sparks and magnitude of caffeine releasable sarcoplasmic reticulum (SR) Ca2+ content. In contrast, high extracellular Ca2+ and ryanodine increased the frequency, full width at half maximum (FWHM) and full duration at half maximum (FDHM) of spontaneous Ca2+ sparks. CONCLUSIONS For the first time, spontaneous Ca2+ sparks were detected in hiPSC-CMs. The Ca2+ sparks are predominately triggered by L-type Ca2+ channels mediated Ca2+ influx, which is comparable to sparks detected in adult ventricular myocytes in which cardiac E-C coupling was governed by a Ca2+-induced Ca2+ release (CICR) mechanism. However, focal repetitive sparks originated from the same intracellular organelle could reflect an immature status of the hiPSC-CMs.", "link"=>"http://www.mendeley.com/research/identification-characterization-calcium-sparks-cardiomyocytes-derived-human-induced-pluripotent-stem-3", "reader_count"=>45, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>8, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>18, "Other"=>2, "Student > Master"=>7, "Student > Bachelor"=>4, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>8, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>18, "Other"=>2, "Student > Master"=>7, "Student > Bachelor"=>4, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>4, "Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>26, "Medicine and Dentistry"=>7, "Physics and Astronomy"=>1, "Computer Science"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>4}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>7}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>26}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"Japan"=>1, "Portugal"=>1, "Russia"=>1, "Spain"=>2}, "group_count"=>3}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/491799"], "description"=>"<p>(A) Immunofluorescent staining of hiPSC colonies with antibodies against Oct-4, SSEA-4, TRA-1-60 and TRA-1-81. (B) The hiPSC-CMs differentia4ed from above hiPSC line. (Ba) The phase-contrast light micrograph images of a V-CM cluster. (Bb and Bc) Immunofluorescent staining hiPSC-CMs with antibodies against alpha-actinin and beta-MHC, respectively. Nuclei were stained with DAPI. (C) Action potential traces of ventricular-, atrial- and nodal-like CMs derived from hiPSCs. (D) Response of a ventricular-like hiPSC-CM to ISO recorded with patch-clamp. Abbreviations: ISO, isoproterenol.</p>", "links"=>[], "tags"=>["hipscs", "hipsc-derived"], "article_id"=>162317, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Guang Qin Zhang", "Heming Wei", "Jun Lu", "Philip Wong", "Winston Shim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055266.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Characterization_of_hiPSCs_and_hiPSC_derived_CMs_/162317", "title"=>"Characterization of hiPSCs and hiPSC-derived CMs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:38:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/492273"], "description"=>"<p>(A) Representative line scan (X-T) images of spontaneous Ca<sup>2+</sup> sparks (top) and intensity-time profiles of typical sparks at positions denoted by white arrows (bottom) before and after the application of nifedipine. The average values of Ca<sup>2+</sup> sparks for frequency (B), F/F<sub>0</sub> (C), FDHM (D) and FWHM (E) before (n<sub>spark</sub> = 213) and after (n<sub>spark</sub> = 128) addition of nifedipine. (F) The line-scan images of caffeine-induced Ca<sup>2+</sup> transients and (G) the corresponding F/F<sub>0</sub> profiles before and after the application of nifedipine. n<sub>cell</sub> = 12. *<i>P</i><0.05 vs. control. Abbreviations: F/F<sub>0</sub>, fluorescence (F) normalized to baseline fluorescence (F<sub>0</sub>); FDHM, full duration at half maximum; FWHM, full width at half maximum; s, seconds.</p>", "links"=>[], "tags"=>["nifedipine", "spontaneous", "sparks", "sr", "loads"], "article_id"=>162793, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Guang Qin Zhang", "Heming Wei", "Jun Lu", "Philip Wong", "Winston Shim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055266.g005", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_nifedipine_on_spontaneous_Ca_2_sparks_and_SR_Ca_2_loads_in_hiPSC_CMs_/162793", "title"=>"Effects of nifedipine on spontaneous Ca<sup>2+</sup> sparks and SR Ca<sup>2+</sup> loads in hiPSC-CMs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:46:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/492368"], "description"=>"<p>(A) Representative line-scan (X-T) images of spontaneous Ca<sup>2+</sup> sparks (top) and the corresponding intensity-time profiles of typical sparks (bottom) before and after the application of 5 mM CaCl<sub>2</sub>. (B) The frequency of Ca<sup>2+</sup> sparks. (C), (D) and (E) show the histograms for F/F<sub>0</sub>, FDHM and FWHM of Ca<sup>2+</sup> sparks before (n<sub>spark</sub> = 143) and after (n<sub>spark</sub> = 318) application of 5 mM CaCl<sub>2</sub>, respectively. n<sub>cell</sub> = 10. *<i>P</i><0.05 vs. control. Abbreviations: F/F<sub>0</sub>, fluorescence (F) normalized to baseline fluorescence (F<sub>0</sub>); FDHM, full duration at half maximum; FWHM, full width at half maximum.</p>", "links"=>[], "tags"=>["spontaneous", "sparks"], "article_id"=>162887, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Guang Qin Zhang", "Heming Wei", "Jun Lu", "Philip Wong", "Winston Shim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055266.g006", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_CaCl_2_on_spontaneous_Ca_2_sparks_in_hiPSC_CMs_/162887", "title"=>"Effects of CaCl<sub>2</sub> on spontaneous Ca<sup>2+</sup> sparks in hiPSC-CMs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:48:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/483585", "https://ndownloader.figshare.com/files/483589", "https://ndownloader.figshare.com/files/483592", "https://ndownloader.figshare.com/files/483595", "https://ndownloader.figshare.com/files/483598", "https://ndownloader.figshare.com/files/483603", "https://ndownloader.figshare.com/files/483606", "https://ndownloader.figshare.com/files/483607"], "description"=>"<div><h3>Introduction</h3><p>Ca<sup>2+</sup> spark constitutes the elementary units of cardiac excitation-contraction (E-C) coupling in mature cardiomyocytes. Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes are known to have electrophysiological properties similar to mature adult cardiomyocytes. However, it is unclear if they share similar calcium handling property. We hypothesized that Ca<sup>2+</sup> sparks in human induced pluripotent stem cell (hiPSCs)-derived cardiomyocytes (hiPSC-CMs) may display unique structural and functional properties than mature adult cardiomyocytes.</p> <h3>Methods and results</h3><p>Ca<sup>2+</sup> sparks in hiPSC-CMs were recorded with Ca<sup>2+</sup> imaging assay with confocal laser scanning microscopy. Those sparks were stochastic with a tendency of repetitive occurrence at the same site. Nevertheless, the spatial-temporal properties of Ca<sup>2+</sup> spark were analogous to that of adult CMs. Inhibition of L-type Ca<sup>2+</sup> channels by nifedipine caused a 61% reduction in calcium spark frequency without affecting amplitude of those sparks and magnitude of caffeine releasable sarcoplasmic reticulum (SR) Ca<sup>2+</sup> content. In contrast, high extracellular Ca<sup>2+</sup> and ryanodine increased the frequency, full width at half maximum (FWHM) and full duration at half maximum (FDHM) of spontaneous Ca<sup>2+</sup> sparks.</p> <h3>Conclusions</h3><p>For the first time<b>,</b> spontaneous Ca<sup>2+</sup> sparks were detected in hiPSC-CMs. The Ca<sup>2+</sup> sparks are predominately triggered by L-type Ca<sup>2+</sup> channels mediated Ca<sup>2+</sup> influx, which is comparable to sparks detected in adult ventricular myocytes in which cardiac E-C coupling was governed by a Ca<sup>2+</sup>-induced Ca<sup>2+</sup> release (CICR) mechanism. However, focal repetitive sparks originated from the same intracellular organelle could reflect an immature status of the hiPSC-CMs.</p> </div>", "links"=>[], "tags"=>["characterization", "calcium", "sparks", "cardiomyocytes", "derived", "induced", "pluripotent", "cells"], "article_id"=>156163, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Guang Qin Zhang", "Heming Wei", "Jun Lu", "Philip Wong", "Winston Shim"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0055266.s001", "https://dx.doi.org/10.1371/journal.pone.0055266.s002", "https://dx.doi.org/10.1371/journal.pone.0055266.s003", "https://dx.doi.org/10.1371/journal.pone.0055266.s004", "https://dx.doi.org/10.1371/journal.pone.0055266.s005", "https://dx.doi.org/10.1371/journal.pone.0055266.s006", "https://dx.doi.org/10.1371/journal.pone.0055266.s007", "https://dx.doi.org/10.1371/journal.pone.0055266.s008"], "stats"=>{"downloads"=>30, "page_views"=>42, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Identification_and_Characterization_of_Calcium_Sparks_in_Cardiomyocytes_Derived_from_Human_Induced_Pluripotent_Stem_Cells__/156163", "title"=>"Identification and Characterization of Calcium Sparks in Cardiomyocytes Derived from Human Induced Pluripotent Stem Cells", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-02-07 01:42:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/492166"], "description"=>"<p>(A) Two representative Ca<sup>2+</sup> sparks (a and b) and an overlay of 160 original Ca<sup>2+</sup> sparks (c) were obtained from the line-scan (X-T) images. (B) The three-dimensional surface plot of the Ca<sup>2+</sup> spark in panel A. (C) The spatial width of Ca<sup>2+</sup> sparks. (D) The duration of Ca<sup>2+</sup> sparks. (E–G) show the distributions of Ca<sup>2+</sup> sparks for F/F<sub>0</sub>, FDHM and FWHM, respectively. n<sub>cell</sub> = 17, n<sub>spark</sub> = 325. *<i>P</i><0.05 vs. control. Abbreviations: F/F<sub>0</sub>, fluorescence (F) normalized to baseline fluorescence (F<sub>0</sub>); FDHM, full duration at half maximum; FWHM, full width at half maximum.</p>", "links"=>[], "tags"=>["characteristics", "spontaneous", "sparks"], "article_id"=>162685, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Guang Qin Zhang", "Heming Wei", "Jun Lu", "Philip Wong", "Winston Shim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055266.g004", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_characteristics_of_spontaneous_Ca_2_sparks_in_hiPSC_CMs_/162685", "title"=>"The characteristics of spontaneous Ca<sup>2+</sup> sparks in hiPSC-CMs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:44:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/492033"], "description"=>"<p>(A) Confocal images of spontaneous sparks were recorded in X-Y scanning mode. Representative spontaneous Ca<sup>2+</sup> sparks occurred at the same site. (B) Two representative line scan (X-T mode) images of Ca<sup>2+</sup> sparks obtained from red line in panel A at different times (top) and the intensity-time profiles of Ca<sup>2+</sup> sparks at sites indicated by white arrows (bottom). Abbreviations: F/F<sub>0</sub>, fluorescence (F) normalized to baseline fluorescence (F<sub>0</sub>).</p>", "links"=>[], "tags"=>["sparks"], "article_id"=>162553, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Guang Qin Zhang", "Heming Wei", "Jun Lu", "Philip Wong", "Winston Shim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055266.g003", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Spontaneous_Ca_2_sparks_in_hiPSC_CMs_/162553", "title"=>"Spontaneous Ca<sup>2+</sup> sparks in hiPSC-CMs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:42:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/491929"], "description"=>"<p>(A) Representative frame-scan (X-Y mode) images of spontaneous Ca<sup>2+</sup> transients (a and b). (B) A typical line scan (X-T mode) image of spontaneous Ca<sup>2+</sup> transients obtained from white line in panel Aa and (C) the corresponding amplitudes (F/F<sub>0</sub>) of Ca<sup>2+</sup> transients (n = 16). (D) A representative transverse line scan (X-T mode) image obtained from green line in panel Aa (a) and the corresponding intensity profiles (b) of Ca<sup>2+</sup> transients. Abbreviations: F/F<sub>0</sub>, fluorescence (F) normalized to baseline fluorescence (F<sub>0</sub>); s, seconds.</p>", "links"=>[], "tags"=>["transients"], "article_id"=>162451, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Guang Qin Zhang", "Heming Wei", "Jun Lu", "Philip Wong", "Winston Shim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055266.g002", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Spontaneous_Ca_2_transients_in_hiPSC_CMs_/162451", "title"=>"Spontaneous Ca<sup>2+</sup> transients in hiPSC-CMs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:40:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/492463"], "description"=>"<p>(A) Representative line-scan (X-T) images of spontaneous Ca<sup>2+</sup> sparks (top) and the corresponding intensity-time profiles of typical sparks (bottom) before and after the application of ryanodine. (B–E) show the mean values for frequency, F/F<sub>0</sub>, FDHM and FWHM of Ca<sup>2+</sup> sparks before (n<sub>spark</sub> = 163) and after (n<sub>spark</sub> = 347) application of ryanodine, respectively. n<sub>cell</sub> = 11. *<i>P</i><0.05 vs. control. Abbreviations: F/F<sub>0</sub>, fluorescence (F) normalized to baseline fluorescence (F<sub>0</sub>); FDHM, full duration at half maximum; FWHM, full width at half maximum.</p>", "links"=>[], "tags"=>["ryanodine", "spontaneous", "sparks"], "article_id"=>162985, "categories"=>["Physiology", "Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Guang Qin Zhang", "Heming Wei", "Jun Lu", "Philip Wong", "Winston Shim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055266.g007", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_ryanodine_on_spontaneous_Ca_2_sparks_in_hiPSC_CMs_/162985", "title"=>"Effects of ryanodine on spontaneous Ca<sup>2+</sup> sparks in hiPSC-CMs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:49:45"}

PMC Usage Stats | Further Information

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Relative Metric

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