Hydrogel microsphere stem cell encapsulation enhances cardiomyocyte differentiation and functionality in scalable suspension system
A reliable suspension-based platform for scaling engineered cardiac tissue (ECT) production from human induced pluripotent stem cells (hiPSCs) is crucial for regenerative therapies. Here, we compared the production and functionality of ECTs formed using our scaffold-based, engineered tissue microsph...
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| Format: | Article |
| Language: | English |
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KeAi Communications Co., Ltd.
2025-01-01
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| Series: | Bioactive Materials |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2452199X24003815 |
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| author | Mohammadjafar Hashemi Ferdous B. Finklea Hanna Hammons Yuan Tian Nathan Young Emma Kim Caroline Halloin Wiebke Triebert Robert Zweigerdt Amit Kumar Mitra Elizabeth A. Lipke |
| author_facet | Mohammadjafar Hashemi Ferdous B. Finklea Hanna Hammons Yuan Tian Nathan Young Emma Kim Caroline Halloin Wiebke Triebert Robert Zweigerdt Amit Kumar Mitra Elizabeth A. Lipke |
| author_sort | Mohammadjafar Hashemi |
| collection | DOAJ |
| description | A reliable suspension-based platform for scaling engineered cardiac tissue (ECT) production from human induced pluripotent stem cells (hiPSCs) is crucial for regenerative therapies. Here, we compared the production and functionality of ECTs formed using our scaffold-based, engineered tissue microsphere differentiation approach with those formed using the prevalent scaffold-free aggregate platform. We utilized a microfluidic system for the rapid (1 million cells/min), high density (30, 40, 60 million cells/ml) encapsulation of hiPSCs within PEG-fibrinogen hydrogel microspheres. HiPSC-laden microspheres and aggregates underwent suspension-based cardiac differentiation in chemically defined media. In comparison to aggregates, microspheres maintained consistent size and shape initially, over time, and within and between batches. Initial size and shape coefficients of variation for microspheres were eight and three times lower, respectively, compared to aggregates. On day 10, microsphere cardiomyocyte (CM) content was 27 % higher and the number of CMs per initial hiPSC was 250 % higher than in aggregates. Contraction and relaxation velocities of microspheres were four and nine times higher than those of aggregates, respectively. Microsphere contractile functionality also improved with culture time, whereas aggregate functionality remained unchanged. Additionally, microspheres displayed improved β-adrenergic signaling responsiveness and uniform calcium transient propagation. Transcriptomic analysis revealed that while both microspheres and aggregates demonstrated similar gene regulation patterns associated with cardiomyocyte differentiation, heart development, cardiac muscle contraction, and sarcomere organization, the microspheres exhibited more pronounced transcriptional changes over time. Taken together, these results highlight the capability of the microsphere platform for scaling up biomanufacturing of ECTs in a suspension-based culture platform. |
| format | Article |
| id | doaj-art-52fce42f65614dd89c7a6701b6a9ef7b |
| institution | Kabale University |
| issn | 2452-199X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Bioactive Materials |
| spelling | doaj-art-52fce42f65614dd89c7a6701b6a9ef7b2024-11-25T04:41:30ZengKeAi Communications Co., Ltd.Bioactive Materials2452-199X2025-01-0143423440Hydrogel microsphere stem cell encapsulation enhances cardiomyocyte differentiation and functionality in scalable suspension systemMohammadjafar Hashemi0Ferdous B. Finklea1Hanna Hammons2Yuan Tian3Nathan Young4Emma Kim5Caroline Halloin6Wiebke Triebert7Robert Zweigerdt8Amit Kumar Mitra9Elizabeth A. Lipke10Department of Chemical Engineering, Auburn University, Auburn, AL, United StatesDepartment of Chemical Engineering, Auburn University, Auburn, AL, United StatesDepartment of Chemical Engineering, Auburn University, Auburn, AL, United StatesDepartment of Chemical Engineering, Auburn University, Auburn, AL, United StatesDepartment of Chemical Engineering, Auburn University, Auburn, AL, United StatesDepartment of Chemical Engineering, Auburn University, Auburn, AL, United StatesLeibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hanover, GermanyLeibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hanover, GermanyLeibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hanover, GermanyDepartment of Drug Discovery and Development, Harrison College of Pharmacy, Auburn University, Auburn, AL, United StatesDepartment of Chemical Engineering, Auburn University, Auburn, AL, United States; Corresponding author.A reliable suspension-based platform for scaling engineered cardiac tissue (ECT) production from human induced pluripotent stem cells (hiPSCs) is crucial for regenerative therapies. Here, we compared the production and functionality of ECTs formed using our scaffold-based, engineered tissue microsphere differentiation approach with those formed using the prevalent scaffold-free aggregate platform. We utilized a microfluidic system for the rapid (1 million cells/min), high density (30, 40, 60 million cells/ml) encapsulation of hiPSCs within PEG-fibrinogen hydrogel microspheres. HiPSC-laden microspheres and aggregates underwent suspension-based cardiac differentiation in chemically defined media. In comparison to aggregates, microspheres maintained consistent size and shape initially, over time, and within and between batches. Initial size and shape coefficients of variation for microspheres were eight and three times lower, respectively, compared to aggregates. On day 10, microsphere cardiomyocyte (CM) content was 27 % higher and the number of CMs per initial hiPSC was 250 % higher than in aggregates. Contraction and relaxation velocities of microspheres were four and nine times higher than those of aggregates, respectively. Microsphere contractile functionality also improved with culture time, whereas aggregate functionality remained unchanged. Additionally, microspheres displayed improved β-adrenergic signaling responsiveness and uniform calcium transient propagation. Transcriptomic analysis revealed that while both microspheres and aggregates demonstrated similar gene regulation patterns associated with cardiomyocyte differentiation, heart development, cardiac muscle contraction, and sarcomere organization, the microspheres exhibited more pronounced transcriptional changes over time. Taken together, these results highlight the capability of the microsphere platform for scaling up biomanufacturing of ECTs in a suspension-based culture platform.http://www.sciencedirect.com/science/article/pii/S2452199X24003815Human induced pluripotent stem cellsSuspension cultureScale-upChemically defined mediaAggregate formationPEG-fibrinogen |
| spellingShingle | Mohammadjafar Hashemi Ferdous B. Finklea Hanna Hammons Yuan Tian Nathan Young Emma Kim Caroline Halloin Wiebke Triebert Robert Zweigerdt Amit Kumar Mitra Elizabeth A. Lipke Hydrogel microsphere stem cell encapsulation enhances cardiomyocyte differentiation and functionality in scalable suspension system Bioactive Materials Human induced pluripotent stem cells Suspension culture Scale-up Chemically defined media Aggregate formation PEG-fibrinogen |
| title | Hydrogel microsphere stem cell encapsulation enhances cardiomyocyte differentiation and functionality in scalable suspension system |
| title_full | Hydrogel microsphere stem cell encapsulation enhances cardiomyocyte differentiation and functionality in scalable suspension system |
| title_fullStr | Hydrogel microsphere stem cell encapsulation enhances cardiomyocyte differentiation and functionality in scalable suspension system |
| title_full_unstemmed | Hydrogel microsphere stem cell encapsulation enhances cardiomyocyte differentiation and functionality in scalable suspension system |
| title_short | Hydrogel microsphere stem cell encapsulation enhances cardiomyocyte differentiation and functionality in scalable suspension system |
| title_sort | hydrogel microsphere stem cell encapsulation enhances cardiomyocyte differentiation and functionality in scalable suspension system |
| topic | Human induced pluripotent stem cells Suspension culture Scale-up Chemically defined media Aggregate formation PEG-fibrinogen |
| url | http://www.sciencedirect.com/science/article/pii/S2452199X24003815 |
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