A systemized approach to investigate Ca2+ synchronization in clusters of human induced pluripotent stem-cell derived cardiomyocytes
Induced pluripotent stem cell-derived cardiomyocytes (IPS-CM) are considered by many to be the cornerstone of future approaches to repair the diseased heart. However, current methods for producing IPS-CM typically yield highly variable populations with low batch-to-batch reproducibility. The underly...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2016-01-01
|
| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fcell.2015.00089/full |
| _version_ | 1811271832274206720 |
|---|---|
| author | Aled R Jones David H Edwards Michael J Cummins Alan J Williams Christopher H George |
| author_facet | Aled R Jones David H Edwards Michael J Cummins Alan J Williams Christopher H George |
| author_sort | Aled R Jones |
| collection | DOAJ |
| description | Induced pluripotent stem cell-derived cardiomyocytes (IPS-CM) are considered by many to be the cornerstone of future approaches to repair the diseased heart. However, current methods for producing IPS-CM typically yield highly variable populations with low batch-to-batch reproducibility. The underlying reasons for this are not fully understood. Here we report on a systematized approach to investigate the effect of maturation in embryoid bodies (EB) versus ‘on plate’ culture on spontaneous activity and regional Ca2+ synchronization in IPS-CM clusters. A detailed analysis of the temporal and spatial organization of Ca2+ spikes in IPS-CM clusters revealed that the disaggregation of EBs between 0.5 and 2 weeks produced IPS-CM characterized by spontaneous beating and high levels of regional Ca2+ synchronization. These phenomena were typically absent in IPS-CM obtained from older EBs (> 2 weeks). The maintenance of all spontaneously active IPS-CM clusters under ‘on plate’ culture conditions promoted the progressive reduction in regional Ca2+ synchronization and the loss of spontaneous Ca2+ spiking. Raising the extracellular [Ca2+] surrounding these quiescent IPS-CM clusters from approximately 0.4 to 1.8 mM unmasked discrete behaviours typified by either a) long-lasting Ca2+ elevation that returned to baseline or b) persistent, large-amplitude Ca2+ oscillations around an increased cytoplasmic [Ca2+]. The different responses of IPS-CM to elevated extracellular [Ca2+] could be traced back to their routes of derivation. The data point to the possibility of predictably influencing IPS-CM phenotype and response to external activation via defined interventions at early stages in their maturation. |
| first_indexed | 2024-04-12T22:29:19Z |
| format | Article |
| id | doaj.art-490b434384694ac884dcb1ee1fae6054 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-04-12T22:29:19Z |
| publishDate | 2016-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-490b434384694ac884dcb1ee1fae60542022-12-22T03:14:02ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2016-01-01310.3389/fcell.2015.00089159341A systemized approach to investigate Ca2+ synchronization in clusters of human induced pluripotent stem-cell derived cardiomyocytesAled R Jones0David H Edwards1Michael J Cummins2Alan J Williams3Christopher H George4Cardiff UniversityCardiff UniversityCardiff UniversityCardiff UniversityCardiff UniversityInduced pluripotent stem cell-derived cardiomyocytes (IPS-CM) are considered by many to be the cornerstone of future approaches to repair the diseased heart. However, current methods for producing IPS-CM typically yield highly variable populations with low batch-to-batch reproducibility. The underlying reasons for this are not fully understood. Here we report on a systematized approach to investigate the effect of maturation in embryoid bodies (EB) versus ‘on plate’ culture on spontaneous activity and regional Ca2+ synchronization in IPS-CM clusters. A detailed analysis of the temporal and spatial organization of Ca2+ spikes in IPS-CM clusters revealed that the disaggregation of EBs between 0.5 and 2 weeks produced IPS-CM characterized by spontaneous beating and high levels of regional Ca2+ synchronization. These phenomena were typically absent in IPS-CM obtained from older EBs (> 2 weeks). The maintenance of all spontaneously active IPS-CM clusters under ‘on plate’ culture conditions promoted the progressive reduction in regional Ca2+ synchronization and the loss of spontaneous Ca2+ spiking. Raising the extracellular [Ca2+] surrounding these quiescent IPS-CM clusters from approximately 0.4 to 1.8 mM unmasked discrete behaviours typified by either a) long-lasting Ca2+ elevation that returned to baseline or b) persistent, large-amplitude Ca2+ oscillations around an increased cytoplasmic [Ca2+]. The different responses of IPS-CM to elevated extracellular [Ca2+] could be traced back to their routes of derivation. The data point to the possibility of predictably influencing IPS-CM phenotype and response to external activation via defined interventions at early stages in their maturation.http://journal.frontiersin.org/Journal/10.3389/fcell.2015.00089/fullInduced Pluripotent Stem CellssynchronizationCalcium signallingvariabilityphenotypeoscillation |
| spellingShingle | Aled R Jones David H Edwards Michael J Cummins Alan J Williams Christopher H George A systemized approach to investigate Ca2+ synchronization in clusters of human induced pluripotent stem-cell derived cardiomyocytes Frontiers in Cell and Developmental Biology Induced Pluripotent Stem Cells synchronization Calcium signalling variability phenotype oscillation |
| title | A systemized approach to investigate Ca2+ synchronization in clusters of human induced pluripotent stem-cell derived cardiomyocytes |
| title_full | A systemized approach to investigate Ca2+ synchronization in clusters of human induced pluripotent stem-cell derived cardiomyocytes |
| title_fullStr | A systemized approach to investigate Ca2+ synchronization in clusters of human induced pluripotent stem-cell derived cardiomyocytes |
| title_full_unstemmed | A systemized approach to investigate Ca2+ synchronization in clusters of human induced pluripotent stem-cell derived cardiomyocytes |
| title_short | A systemized approach to investigate Ca2+ synchronization in clusters of human induced pluripotent stem-cell derived cardiomyocytes |
| title_sort | systemized approach to investigate ca2 synchronization in clusters of human induced pluripotent stem cell derived cardiomyocytes |
| topic | Induced Pluripotent Stem Cells synchronization Calcium signalling variability phenotype oscillation |
| url | http://journal.frontiersin.org/Journal/10.3389/fcell.2015.00089/full |
| work_keys_str_mv | AT aledrjones asystemizedapproachtoinvestigateca2synchronizationinclustersofhumaninducedpluripotentstemcellderivedcardiomyocytes AT davidhedwards asystemizedapproachtoinvestigateca2synchronizationinclustersofhumaninducedpluripotentstemcellderivedcardiomyocytes AT michaeljcummins asystemizedapproachtoinvestigateca2synchronizationinclustersofhumaninducedpluripotentstemcellderivedcardiomyocytes AT alanjwilliams asystemizedapproachtoinvestigateca2synchronizationinclustersofhumaninducedpluripotentstemcellderivedcardiomyocytes AT christopherhgeorge asystemizedapproachtoinvestigateca2synchronizationinclustersofhumaninducedpluripotentstemcellderivedcardiomyocytes AT aledrjones systemizedapproachtoinvestigateca2synchronizationinclustersofhumaninducedpluripotentstemcellderivedcardiomyocytes AT davidhedwards systemizedapproachtoinvestigateca2synchronizationinclustersofhumaninducedpluripotentstemcellderivedcardiomyocytes AT michaeljcummins systemizedapproachtoinvestigateca2synchronizationinclustersofhumaninducedpluripotentstemcellderivedcardiomyocytes AT alanjwilliams systemizedapproachtoinvestigateca2synchronizationinclustersofhumaninducedpluripotentstemcellderivedcardiomyocytes AT christopherhgeorge systemizedapproachtoinvestigateca2synchronizationinclustersofhumaninducedpluripotentstemcellderivedcardiomyocytes |