A systemic evaluation of cardiac differentiation from mRNA reprogrammed human induced pluripotent stem cells.
Genetically unmodified cardiomyocytes mandated for cardiac regenerative therapy is conceivable by "foot-print free" reprogramming of somatic cells to induced pluripotent stem cells (iPSC). In this study, we report generation of foot-print free hiPSC through messenger RNA (mRNA) based repro...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4113436?pdf=render |
| _version_ | 1818314648328339456 |
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| author | Ashish Mehta Vinod Verma Manasi Nandihalli Chrishan J A Ramachandra Glen L Sequiera Yuliansa Sudibyo Yingying Chung William Sun Winston Shim |
| author_facet | Ashish Mehta Vinod Verma Manasi Nandihalli Chrishan J A Ramachandra Glen L Sequiera Yuliansa Sudibyo Yingying Chung William Sun Winston Shim |
| author_sort | Ashish Mehta |
| collection | DOAJ |
| description | Genetically unmodified cardiomyocytes mandated for cardiac regenerative therapy is conceivable by "foot-print free" reprogramming of somatic cells to induced pluripotent stem cells (iPSC). In this study, we report generation of foot-print free hiPSC through messenger RNA (mRNA) based reprograming. Subsequently, we characterize cardiomyocytes derived from these hiPSC using molecular and electrophysiological methods to characterize their applicability for regenerative medicine. Our results demonstrate that mRNA-iPSCs differentiate ontogenetically into cardiomyocytes with increased expression of early commitment markers of mesoderm, cardiac mesoderm, followed by cardiac specific transcriptional and sarcomeric structural and ion channel genes. Furthermore, these cardiomyocytes stained positively for sarcomeric and ion channel proteins. Based on multi-electrode array (MEA) recordings, these mRNA-hiPSC derived cardiomyocytes responded predictably to various pharmacologically active drugs that target adrenergic, sodium, calcium and potassium channels. The cardiomyocytes responded chronotropically to isoproterenol in a dose dependent manner, inotropic activity of nifidipine decreased spontaneous contractions. Moreover, Sotalol and E-4031 prolonged QT intervals, while TTX reduced sodium influx. Our results for the first time show a systemic evaluation based on molecular, structural and functional properties of cardiomyocytes differentiated from mRNA-iPSC. These results, coupled with feasibility of generating patient-specific iPSCs hold great promise for the development of large-scale generation of clinical grade cardiomyocytes for cardiac regenerative medicine. |
| first_indexed | 2024-12-13T08:52:59Z |
| format | Article |
| id | doaj.art-cde1246d4b924048b218f43c7e79e4a5 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T08:52:59Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-cde1246d4b924048b218f43c7e79e4a52022-12-21T23:53:20ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0197e10348510.1371/journal.pone.0103485A systemic evaluation of cardiac differentiation from mRNA reprogrammed human induced pluripotent stem cells.Ashish MehtaVinod VermaManasi NandihalliChrishan J A RamachandraGlen L SequieraYuliansa SudibyoYingying ChungWilliam SunWinston ShimGenetically unmodified cardiomyocytes mandated for cardiac regenerative therapy is conceivable by "foot-print free" reprogramming of somatic cells to induced pluripotent stem cells (iPSC). In this study, we report generation of foot-print free hiPSC through messenger RNA (mRNA) based reprograming. Subsequently, we characterize cardiomyocytes derived from these hiPSC using molecular and electrophysiological methods to characterize their applicability for regenerative medicine. Our results demonstrate that mRNA-iPSCs differentiate ontogenetically into cardiomyocytes with increased expression of early commitment markers of mesoderm, cardiac mesoderm, followed by cardiac specific transcriptional and sarcomeric structural and ion channel genes. Furthermore, these cardiomyocytes stained positively for sarcomeric and ion channel proteins. Based on multi-electrode array (MEA) recordings, these mRNA-hiPSC derived cardiomyocytes responded predictably to various pharmacologically active drugs that target adrenergic, sodium, calcium and potassium channels. The cardiomyocytes responded chronotropically to isoproterenol in a dose dependent manner, inotropic activity of nifidipine decreased spontaneous contractions. Moreover, Sotalol and E-4031 prolonged QT intervals, while TTX reduced sodium influx. Our results for the first time show a systemic evaluation based on molecular, structural and functional properties of cardiomyocytes differentiated from mRNA-iPSC. These results, coupled with feasibility of generating patient-specific iPSCs hold great promise for the development of large-scale generation of clinical grade cardiomyocytes for cardiac regenerative medicine.http://europepmc.org/articles/PMC4113436?pdf=render |
| spellingShingle | Ashish Mehta Vinod Verma Manasi Nandihalli Chrishan J A Ramachandra Glen L Sequiera Yuliansa Sudibyo Yingying Chung William Sun Winston Shim A systemic evaluation of cardiac differentiation from mRNA reprogrammed human induced pluripotent stem cells. PLoS ONE |
| title | A systemic evaluation of cardiac differentiation from mRNA reprogrammed human induced pluripotent stem cells. |
| title_full | A systemic evaluation of cardiac differentiation from mRNA reprogrammed human induced pluripotent stem cells. |
| title_fullStr | A systemic evaluation of cardiac differentiation from mRNA reprogrammed human induced pluripotent stem cells. |
| title_full_unstemmed | A systemic evaluation of cardiac differentiation from mRNA reprogrammed human induced pluripotent stem cells. |
| title_short | A systemic evaluation of cardiac differentiation from mRNA reprogrammed human induced pluripotent stem cells. |
| title_sort | systemic evaluation of cardiac differentiation from mrna reprogrammed human induced pluripotent stem cells |
| url | http://europepmc.org/articles/PMC4113436?pdf=render |
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