Specific induction and long-term maintenance of high purity ventricular cardiomyocytes from human induced pluripotent stem cells.
Currently, cardiomyocyte (CM) differentiation methods require a purification step after CM induction to ensure the high purity of the cell population. Here we show an improved human CM differentiation protocol with which high-purity ventricular-type CMs can be obtained and maintained without any CM...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2020-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0241287 |
| _version_ | 1818398546552946688 |
|---|---|
| author | Hiroyuki Fukushima Miki Yoshioka Masahide Kawatou Víctor López-Dávila Masafumi Takeda Yasunari Kanda Yuko Sekino Yoshinori Yoshida Jun K Yamashita |
| author_facet | Hiroyuki Fukushima Miki Yoshioka Masahide Kawatou Víctor López-Dávila Masafumi Takeda Yasunari Kanda Yuko Sekino Yoshinori Yoshida Jun K Yamashita |
| author_sort | Hiroyuki Fukushima |
| collection | DOAJ |
| description | Currently, cardiomyocyte (CM) differentiation methods require a purification step after CM induction to ensure the high purity of the cell population. Here we show an improved human CM differentiation protocol with which high-purity ventricular-type CMs can be obtained and maintained without any CM purification process. We induced and collected a mesodermal cell population (platelet-derived growth factor receptor-α (PDGFRα)-positive cells) that can respond to CM differentiation cues, and then stimulated CM differentiation by means of Wnt inhibition. This method reproducibly generated CMs with purities above 95% in several human pluripotent stem cell lines. Furthermore, these CM populations were maintained in culture at such high purity without any further CM purification step for over 200 days. The majority of these CMs (>95%) exhibited a ventricular-like phenotype with a tendency to structural and electrophysiological maturation, including T-tubule-like structure formation and the ability to respond to QT prolongation drugs. This is a simple and valuable method to stably generate CM populations suitable for cardiac toxicology testing, disease modeling and regenerative medicine. |
| first_indexed | 2024-12-14T07:06:30Z |
| format | Article |
| id | doaj.art-3b2ecb5d3fd7405295612bd0a6aef181 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-14T07:06:30Z |
| publishDate | 2020-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-3b2ecb5d3fd7405295612bd0a6aef1812022-12-21T23:12:10ZengPublic Library of Science (PLoS)PLoS ONE1932-62032020-01-011511e024128710.1371/journal.pone.0241287Specific induction and long-term maintenance of high purity ventricular cardiomyocytes from human induced pluripotent stem cells.Hiroyuki FukushimaMiki YoshiokaMasahide KawatouVíctor López-DávilaMasafumi TakedaYasunari KandaYuko SekinoYoshinori YoshidaJun K YamashitaCurrently, cardiomyocyte (CM) differentiation methods require a purification step after CM induction to ensure the high purity of the cell population. Here we show an improved human CM differentiation protocol with which high-purity ventricular-type CMs can be obtained and maintained without any CM purification process. We induced and collected a mesodermal cell population (platelet-derived growth factor receptor-α (PDGFRα)-positive cells) that can respond to CM differentiation cues, and then stimulated CM differentiation by means of Wnt inhibition. This method reproducibly generated CMs with purities above 95% in several human pluripotent stem cell lines. Furthermore, these CM populations were maintained in culture at such high purity without any further CM purification step for over 200 days. The majority of these CMs (>95%) exhibited a ventricular-like phenotype with a tendency to structural and electrophysiological maturation, including T-tubule-like structure formation and the ability to respond to QT prolongation drugs. This is a simple and valuable method to stably generate CM populations suitable for cardiac toxicology testing, disease modeling and regenerative medicine.https://doi.org/10.1371/journal.pone.0241287 |
| spellingShingle | Hiroyuki Fukushima Miki Yoshioka Masahide Kawatou Víctor López-Dávila Masafumi Takeda Yasunari Kanda Yuko Sekino Yoshinori Yoshida Jun K Yamashita Specific induction and long-term maintenance of high purity ventricular cardiomyocytes from human induced pluripotent stem cells. PLoS ONE |
| title | Specific induction and long-term maintenance of high purity ventricular cardiomyocytes from human induced pluripotent stem cells. |
| title_full | Specific induction and long-term maintenance of high purity ventricular cardiomyocytes from human induced pluripotent stem cells. |
| title_fullStr | Specific induction and long-term maintenance of high purity ventricular cardiomyocytes from human induced pluripotent stem cells. |
| title_full_unstemmed | Specific induction and long-term maintenance of high purity ventricular cardiomyocytes from human induced pluripotent stem cells. |
| title_short | Specific induction and long-term maintenance of high purity ventricular cardiomyocytes from human induced pluripotent stem cells. |
| title_sort | specific induction and long term maintenance of high purity ventricular cardiomyocytes from human induced pluripotent stem cells |
| url | https://doi.org/10.1371/journal.pone.0241287 |
| work_keys_str_mv | AT hiroyukifukushima specificinductionandlongtermmaintenanceofhighpurityventricularcardiomyocytesfromhumaninducedpluripotentstemcells AT mikiyoshioka specificinductionandlongtermmaintenanceofhighpurityventricularcardiomyocytesfromhumaninducedpluripotentstemcells AT masahidekawatou specificinductionandlongtermmaintenanceofhighpurityventricularcardiomyocytesfromhumaninducedpluripotentstemcells AT victorlopezdavila specificinductionandlongtermmaintenanceofhighpurityventricularcardiomyocytesfromhumaninducedpluripotentstemcells AT masafumitakeda specificinductionandlongtermmaintenanceofhighpurityventricularcardiomyocytesfromhumaninducedpluripotentstemcells AT yasunarikanda specificinductionandlongtermmaintenanceofhighpurityventricularcardiomyocytesfromhumaninducedpluripotentstemcells AT yukosekino specificinductionandlongtermmaintenanceofhighpurityventricularcardiomyocytesfromhumaninducedpluripotentstemcells AT yoshinoriyoshida specificinductionandlongtermmaintenanceofhighpurityventricularcardiomyocytesfromhumaninducedpluripotentstemcells AT junkyamashita specificinductionandlongtermmaintenanceofhighpurityventricularcardiomyocytesfromhumaninducedpluripotentstemcells |