Tissue-engineered human embryonic stem cell-containing cardiac patches: evaluating recellularization of decellularized matrix
Autor: | Rafael Serafim Pinto, Gustavo Monnerat, Gilberto Weissmüller, Dilza Campos, Luiz C. Sampaio, Antonio Carlos Campos de Carvalho, Camila Hochman-Mendez, Doris A. Taylor, Adriana Bastos Carvalho, Bernardo Jorge da Silva Mendes, Gustavo Miranda Rocha |
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Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
Confocal
Biomedical Engineering Medicine (miscellaneous) Connexin cardiomyocytes mechanical properties Sarcomere Biomaterials Extracellular matrix lcsh:Biochemistry 03 medical and health sciences 0302 clinical medicine Tissue engineering medicine lcsh:QD415-436 030304 developmental biology Basement membrane 0303 health sciences Decellularization Chemistry Embryonic stem cell Cell biology medicine.anatomical_structure recellularization efficiency Original Article 030217 neurology & neurosurgery Decellularized extracellular matrix |
Zdroj: | Journal of Tissue Engineering, Vol 11 (2020) Journal of Tissue Engineering |
ISSN: | 2041-7314 |
Popis: | Decellularized cardiac extracellular matrix scaffolds with preserved composition and architecture can be used in tissue engineering to reproduce the complex cardiac extracellular matrix. However, evaluating the extent of cardiomyocyte repopulation of decellularized cardiac extracellular matrix scaffolds after recellularization attempts is challenging. Here, we describe a unique combination of biochemical, biomechanical, histological, and physiological parameters for quantifying recellularization efficiency of tissue-engineered cardiac patches compared with native cardiac tissue. Human embryonic stem cell-derived cardiomyocytes were seeded into rat heart atrial and ventricular decellularized cardiac extracellular matrix patches. Confocal and atomic force microscopy showed cell integration within the extracellular matrix basement membrane that was accompanied by restoration of native cardiac tissue passive mechanical properties. Multi-electrode array and immunostaining (connexin 43) were used to determine synchronous field potentials with electrical coupling. Myoglobin content (~60%) and sarcomere length measurement (>45% vs 2D culture) were used to evaluate cardiomyocyte maturation of integrated cells. The combination of these techniques allowed us to demonstrate that as cellularization efficiency improves, cardiomyocytes mature and synchronize electrical activity, and tissue mechanical/biochemical properties improve toward those of native tissue. |
Databáze: | OpenAIRE |
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