Timely delivery of cardiac mmRNAs in microfluidics enhances cardiogenic programming of human pluripotent stem cells
In the last two decades lab-on-chip models, specifically heart-on-chip, have been developed as promising technologies for recapitulating physiological environments suitable for studies of drug and environmental effects on either human physiological or patho-physiological conditions. Most human heart...
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| Format: | Article |
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Frontiers Media S.A.
2022-08-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2022.871867/full |
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| author | Anna Contato Anna Contato Onelia Gagliano Onelia Gagliano Michael Magnussen Monica Giomo Nicola Elvassore Nicola Elvassore Nicola Elvassore |
| author_facet | Anna Contato Anna Contato Onelia Gagliano Onelia Gagliano Michael Magnussen Monica Giomo Nicola Elvassore Nicola Elvassore Nicola Elvassore |
| author_sort | Anna Contato |
| collection | DOAJ |
| description | In the last two decades lab-on-chip models, specifically heart-on-chip, have been developed as promising technologies for recapitulating physiological environments suitable for studies of drug and environmental effects on either human physiological or patho-physiological conditions. Most human heart-on-chip systems are based on integration and adaptation of terminally differentiated cells within microfluidic context. This process requires prolonged procedures, multiple steps, and is associated with an intrinsic variability of cardiac differentiation. In this view, we developed a method for cardiac differentiation-on-a-chip based on combining the stage-specific regulation of Wnt/β-catenin signaling with the forced expression of transcription factors (TFs) that timely recapitulate hallmarks of the cardiac development. We performed the overall cardiac differentiation from human pluripotent stem cells (hPSCs) to cardiomyocytes (CMs) within a microfluidic environment. Sequential forced expression of cardiac TFs was achieved by a sequential mmRNAs delivery of first MESP1, GATA4 followed by GATA4, NKX2.5, MEF2C, TBX3, and TBX5. We showed that this optimized protocol led to a robust and reproducible approach to obtain a cost-effective hiPSC-derived heart-on-chip. The results showed higher distribution of cTNT positive CMs along the channel and a higher expression of functional cardiac markers (TNNT2 and MYH7). The combination of stage-specific regulation of Wnt/β-catenin signaling with mmRNAs encoding cardiac transcription factors will be suitable to obtain heart-on-chip model in a cost-effective manner, enabling to perform combinatorial, multiparametric, parallelized and high-throughput experiments on functional cardiomyocytes. |
| first_indexed | 2024-12-10T20:19:29Z |
| format | Article |
| id | doaj.art-a6aa4a25182545b2a21d24cf7b0fad59 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-12-10T20:19:29Z |
| publishDate | 2022-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-a6aa4a25182545b2a21d24cf7b0fad592022-12-22T01:35:05ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-08-011010.3389/fbioe.2022.871867871867Timely delivery of cardiac mmRNAs in microfluidics enhances cardiogenic programming of human pluripotent stem cellsAnna Contato0Anna Contato1Onelia Gagliano2Onelia Gagliano3Michael Magnussen4Monica Giomo5Nicola Elvassore6Nicola Elvassore7Nicola Elvassore8Department of Industrial Engineering, University of Padova, Padova, ItalyVeneto Institute of Molecular Medicine, Padova, ItalyDepartment of Industrial Engineering, University of Padova, Padova, ItalyVeneto Institute of Molecular Medicine, Padova, ItalyGOSICH Zayed Centre for Research Into Rare Disease in Children, University College London, London, United KingdomDepartment of Industrial Engineering, University of Padova, Padova, ItalyDepartment of Industrial Engineering, University of Padova, Padova, ItalyVeneto Institute of Molecular Medicine, Padova, ItalyGOSICH Zayed Centre for Research Into Rare Disease in Children, University College London, London, United KingdomIn the last two decades lab-on-chip models, specifically heart-on-chip, have been developed as promising technologies for recapitulating physiological environments suitable for studies of drug and environmental effects on either human physiological or patho-physiological conditions. Most human heart-on-chip systems are based on integration and adaptation of terminally differentiated cells within microfluidic context. This process requires prolonged procedures, multiple steps, and is associated with an intrinsic variability of cardiac differentiation. In this view, we developed a method for cardiac differentiation-on-a-chip based on combining the stage-specific regulation of Wnt/β-catenin signaling with the forced expression of transcription factors (TFs) that timely recapitulate hallmarks of the cardiac development. We performed the overall cardiac differentiation from human pluripotent stem cells (hPSCs) to cardiomyocytes (CMs) within a microfluidic environment. Sequential forced expression of cardiac TFs was achieved by a sequential mmRNAs delivery of first MESP1, GATA4 followed by GATA4, NKX2.5, MEF2C, TBX3, and TBX5. We showed that this optimized protocol led to a robust and reproducible approach to obtain a cost-effective hiPSC-derived heart-on-chip. The results showed higher distribution of cTNT positive CMs along the channel and a higher expression of functional cardiac markers (TNNT2 and MYH7). The combination of stage-specific regulation of Wnt/β-catenin signaling with mmRNAs encoding cardiac transcription factors will be suitable to obtain heart-on-chip model in a cost-effective manner, enabling to perform combinatorial, multiparametric, parallelized and high-throughput experiments on functional cardiomyocytes.https://www.frontiersin.org/articles/10.3389/fbioe.2022.871867/fullmmRNAsmicrofluidicsheart-on-chipcardiogenesishPSCs |
| spellingShingle | Anna Contato Anna Contato Onelia Gagliano Onelia Gagliano Michael Magnussen Monica Giomo Nicola Elvassore Nicola Elvassore Nicola Elvassore Timely delivery of cardiac mmRNAs in microfluidics enhances cardiogenic programming of human pluripotent stem cells Frontiers in Bioengineering and Biotechnology mmRNAs microfluidics heart-on-chip cardiogenesis hPSCs |
| title | Timely delivery of cardiac mmRNAs in microfluidics enhances cardiogenic programming of human pluripotent stem cells |
| title_full | Timely delivery of cardiac mmRNAs in microfluidics enhances cardiogenic programming of human pluripotent stem cells |
| title_fullStr | Timely delivery of cardiac mmRNAs in microfluidics enhances cardiogenic programming of human pluripotent stem cells |
| title_full_unstemmed | Timely delivery of cardiac mmRNAs in microfluidics enhances cardiogenic programming of human pluripotent stem cells |
| title_short | Timely delivery of cardiac mmRNAs in microfluidics enhances cardiogenic programming of human pluripotent stem cells |
| title_sort | timely delivery of cardiac mmrnas in microfluidics enhances cardiogenic programming of human pluripotent stem cells |
| topic | mmRNAs microfluidics heart-on-chip cardiogenesis hPSCs |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2022.871867/full |
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