In Vivo Cannulation Methods for Cardiomyocytes Isolation from Heart Disease Models.
Isolation of high quality cardiomyocytes is critically important for achieving successful experiments in many cellular and molecular cardiology studies. Methods for isolating cardiomyocytes from the murine heart generally are time-sensitive and experience-dependent, and often fail to produce high qu...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2016-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4976940?pdf=render |
| _version_ | 1828463553981448192 |
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| author | Zhong Jian Yi-Je Chen Rafael Shimkunas Yuwen Jian Mark Jaradeh Karen Chavez Nipavan Chiamvimonvat Jil C Tardiff Leighton T Izu Robert S Ross Ye Chen-Izu |
| author_facet | Zhong Jian Yi-Je Chen Rafael Shimkunas Yuwen Jian Mark Jaradeh Karen Chavez Nipavan Chiamvimonvat Jil C Tardiff Leighton T Izu Robert S Ross Ye Chen-Izu |
| author_sort | Zhong Jian |
| collection | DOAJ |
| description | Isolation of high quality cardiomyocytes is critically important for achieving successful experiments in many cellular and molecular cardiology studies. Methods for isolating cardiomyocytes from the murine heart generally are time-sensitive and experience-dependent, and often fail to produce high quality cells. Major technical difficulties can be related to the surgical procedures needed to explant the heart and to cannulate the vessel to mount onto the Langendorff system before in vitro reperfusion can begin. During this period, transient hypoxia and ischemia may damage the heart, resulting in low yield and poor quality of cells, especially for heart disease models that have fragile cells. We have developed novel in vivo cannulation methods to minimize hypoxia and ischemia, and fine-tuned the entire protocol to produce high quality ventricular myocytes. The high cell quality has been confirmed using important structural and functional criteria such as morphology, t-tubule structure, action potential morphology, Ca2+ signaling, responsiveness to beta-adrenergic agonist, and ability to have robust contraction under mechanically loaded condition. Together these assessments show the preservation of the cardiac excitation-contraction machinery in cells isolated using this technique. The in vivo cannulation method enables consistent isolation of high-quality cardiomyocytes, even from heart disease models that were notoriously difficult for cell isolation using traditional methods. |
| first_indexed | 2024-12-11T02:56:39Z |
| format | Article |
| id | doaj.art-2310d847ebcf4ff785924dcfc7a34c51 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-11T02:56:39Z |
| publishDate | 2016-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-2310d847ebcf4ff785924dcfc7a34c512022-12-22T01:23:08ZengPublic Library of Science (PLoS)PLoS ONE1932-62032016-01-01118e016060510.1371/journal.pone.0160605In Vivo Cannulation Methods for Cardiomyocytes Isolation from Heart Disease Models.Zhong JianYi-Je ChenRafael ShimkunasYuwen JianMark JaradehKaren ChavezNipavan ChiamvimonvatJil C TardiffLeighton T IzuRobert S RossYe Chen-IzuIsolation of high quality cardiomyocytes is critically important for achieving successful experiments in many cellular and molecular cardiology studies. Methods for isolating cardiomyocytes from the murine heart generally are time-sensitive and experience-dependent, and often fail to produce high quality cells. Major technical difficulties can be related to the surgical procedures needed to explant the heart and to cannulate the vessel to mount onto the Langendorff system before in vitro reperfusion can begin. During this period, transient hypoxia and ischemia may damage the heart, resulting in low yield and poor quality of cells, especially for heart disease models that have fragile cells. We have developed novel in vivo cannulation methods to minimize hypoxia and ischemia, and fine-tuned the entire protocol to produce high quality ventricular myocytes. The high cell quality has been confirmed using important structural and functional criteria such as morphology, t-tubule structure, action potential morphology, Ca2+ signaling, responsiveness to beta-adrenergic agonist, and ability to have robust contraction under mechanically loaded condition. Together these assessments show the preservation of the cardiac excitation-contraction machinery in cells isolated using this technique. The in vivo cannulation method enables consistent isolation of high-quality cardiomyocytes, even from heart disease models that were notoriously difficult for cell isolation using traditional methods.http://europepmc.org/articles/PMC4976940?pdf=render |
| spellingShingle | Zhong Jian Yi-Je Chen Rafael Shimkunas Yuwen Jian Mark Jaradeh Karen Chavez Nipavan Chiamvimonvat Jil C Tardiff Leighton T Izu Robert S Ross Ye Chen-Izu In Vivo Cannulation Methods for Cardiomyocytes Isolation from Heart Disease Models. PLoS ONE |
| title | In Vivo Cannulation Methods for Cardiomyocytes Isolation from Heart Disease Models. |
| title_full | In Vivo Cannulation Methods for Cardiomyocytes Isolation from Heart Disease Models. |
| title_fullStr | In Vivo Cannulation Methods for Cardiomyocytes Isolation from Heart Disease Models. |
| title_full_unstemmed | In Vivo Cannulation Methods for Cardiomyocytes Isolation from Heart Disease Models. |
| title_short | In Vivo Cannulation Methods for Cardiomyocytes Isolation from Heart Disease Models. |
| title_sort | in vivo cannulation methods for cardiomyocytes isolation from heart disease models |
| url | http://europepmc.org/articles/PMC4976940?pdf=render |
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