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10 18, 2022;12 (1): 17409.

Generation and maturation of human iPSC-derived 3D organotypic cardiac microtissues in long-term culture.

Ece Ergir, Jorge Oliver-De La Cruz, Soraia Fernandes, Marco Cassani, Francesco Niro, Daniel Pereira-Sousa, Jan Vrbský, Vladimír Vinarský, Ana Rubina Perestrelo, Doriana Debellis, Natália Vadovičová, Stjepan Uldrijan, Francesca Cavalieri, Stefania Pagliari, Heinz Redl, Peter Ertl, Giancarlo Forte
Author Information
  1. Ece Ergir: Center for Translational Medicine (CTM), International Clinical Research Centre (FNUSA-ICRC), St. Anne's University Hospital, Studentská 812/6, 62500, Brno, Czech Republic.
  2. Jorge Oliver-De La Cruz: Center for Translational Medicine (CTM), International Clinical Research Centre (FNUSA-ICRC), St. Anne's University Hospital, Studentská 812/6, 62500, Brno, Czech Republic.
  3. Soraia Fernandes: Center for Translational Medicine (CTM), International Clinical Research Centre (FNUSA-ICRC), St. Anne's University Hospital, Studentská 812/6, 62500, Brno, Czech Republic.
  4. Marco Cassani: Center for Translational Medicine (CTM), International Clinical Research Centre (FNUSA-ICRC), St. Anne's University Hospital, Studentská 812/6, 62500, Brno, Czech Republic.
  5. Francesco Niro: Center for Translational Medicine (CTM), International Clinical Research Centre (FNUSA-ICRC), St. Anne's University Hospital, Studentská 812/6, 62500, Brno, Czech Republic.
  6. Daniel Pereira-Sousa: Center for Translational Medicine (CTM), International Clinical Research Centre (FNUSA-ICRC), St. Anne's University Hospital, Studentská 812/6, 62500, Brno, Czech Republic.
  7. Jan Vrbský: Center for Translational Medicine (CTM), International Clinical Research Centre (FNUSA-ICRC), St. Anne's University Hospital, Studentská 812/6, 62500, Brno, Czech Republic.
  8. Vladimír Vinarský: Center for Translational Medicine (CTM), International Clinical Research Centre (FNUSA-ICRC), St. Anne's University Hospital, Studentská 812/6, 62500, Brno, Czech Republic.
  9. Ana Rubina Perestrelo: Center for Translational Medicine (CTM), International Clinical Research Centre (FNUSA-ICRC), St. Anne's University Hospital, Studentská 812/6, 62500, Brno, Czech Republic.
  10. Doriana Debellis: Electron Microscopy Facility, Fondazione Istituto Italiano Di Tecnologia, Via Morego 30, 16163, Genoa, Italy.
  11. Natália Vadovičová: Faculty of Medicine, Department of Biomedical Sciences, Masaryk University, 62500, Brno, Czech Republic.
  12. Stjepan Uldrijan: Faculty of Medicine, Department of Biomedical Sciences, Masaryk University, 62500, Brno, Czech Republic. ORCID
  13. Francesca Cavalieri: Department of Chemical Engineering, The University of Melbourne, Parkville, VIC, 3010, Australia.
  14. Stefania Pagliari: Center for Translational Medicine (CTM), International Clinical Research Centre (FNUSA-ICRC), St. Anne's University Hospital, Studentská 812/6, 62500, Brno, Czech Republic.
  15. Heinz Redl: Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center, 1200, Vienna, Austria.
  16. Peter Ertl: Faculty of Technical Chemistry, Institute of Applied Synthetic Chemistry and Institute of Chemical Technologies and Analytics, Vienna University of Technology, 1040, Vienna, Austria.
  17. Giancarlo Forte: Center for Translational Medicine (CTM), International Clinical Research Centre (FNUSA-ICRC), St. Anne's University Hospital, Studentská 812/6, 62500, Brno, Czech Republic. giancarlo.forte@fnusa.cz. ORCID
PMID: 36257968 DOI: 10.1038/s41598-022-22225-w

Abstract

Cardiovascular diseases remain the leading cause of death worldwide; hence there is an increasing focus on developing physiologically relevant in vitro cardiovascular tissue models suitable for studying personalized medicine and pre-clinical tests. Despite recent advances, models that reproduce both tissue complexity and maturation are still limited. We have established a scaffold-free protocol to generate multicellular, beating human cardiac microtissues in vitro from hiPSCs-namely human organotypic cardiac microtissues (hOCMTs)-that show some degree of self-organization and can be cultured for long term. This is achieved by the differentiation of hiPSC in 2D monolayer culture towards cardiovascular lineage, followed by further aggregation on low-attachment culture dishes in 3D. The generated hOCMTs contain multiple cell types that physiologically compose the heart and beat without external stimuli for more than 100 days. We have shown that 3D hOCMTs display improved cardiac specification, survival and metabolic maturation as compared to standard monolayer cardiac differentiation. We also confirmed the functionality of hOCMTs by their response to cardioactive drugs in long-term culture. Furthermore, we demonstrated that they could be used to study chemotherapy-induced cardiotoxicity. Due to showing a tendency for self-organization, cellular heterogeneity, and functionality in our 3D microtissues over extended culture time, we could also confirm these constructs as human cardiac organoids (hCOs). This study could help to develop more physiologically-relevant cardiac tissue models, and represent a powerful platform for future translational research in cardiovascular biology.

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MeSH Term

Humans
Induced Pluripotent Stem Cells
Tissue Engineering
Heart
Cell Differentiation
Cardiovascular Agents
Antineoplastic Agents
Myocytes, Cardiac

Chemicals

Cardiovascular Agents
Antineoplastic Agents
Journal Article Research Support, Non-U.S. Gov't
Article has an altmetric score of 12

Word Cloud

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