Adult Multipotent Cardiac Progenitor-Derived Spheroids: A Reproducible Model of In Vitro Cardiomyocyte Commitment and Specification

Three-dimensional cell culture systems hold great promise for bridging the gap between in vitro cell-based model systems and small animal models to study tissue biology and disease. Among 3D cell culture systems, stem-cell-derived spheroids have attracted significant interest as a strategy to better...

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Published in:	Cells (Basel, Switzerland) Vol. 12; no. 13; p. 1793
Main Authors:	Scalise, Mariangela, Marino, Fabiola, Salerno, Luca, Amato, Nunzia, Quercia, Claudia, Siracusa, Chiara, Filardo, Andrea, Chiefalo, Antonio, Pagano, Loredana, Misdea, Giuseppe, Salerno, Nadia, De Angelis, Antonella, Urbanek, Konrad, Viglietto, Giuseppe, Torella, Daniele, Cianflone, Eleonora
Format:	Journal Article
Language:	English
Published:	Switzerland MDPI AG 01-07-2023 MDPI
Subjects:	3D cell culture Analysis Animal models Animals cardiac spheroids cardiomyocyte differentiation Cardiomyocytes Cell culture Cell Differentiation Cloning Growth factors Health aspects Heart Heart cells Hematopoietic Stem Cells Hypoxia Methods Microenvironments multipotent cardiac progenitors Myocytes, Cardiac Progenitor cells Regenerative medicine Sepharose Spheroids Stem cells Yeast Italy United Kingdom > UK United States > US Germany cardiomyocyte differentiation 3D cell culture cardiac spheroids multipotent cardiac progenitors
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Summary:	Three-dimensional cell culture systems hold great promise for bridging the gap between in vitro cell-based model systems and small animal models to study tissue biology and disease. Among 3D cell culture systems, stem-cell-derived spheroids have attracted significant interest as a strategy to better mimic in vivo conditions. Cardiac stem cell/progenitor (CSC)-derived spheroids (CSs) provide a relevant platform for cardiac regeneration. We compared three different cell culture scaffold-free systems, (i) ultra-low attachment plates, (ii) hanging drops (both requiring a 2D/3D switch), and (iii) agarose micro-molds (entirely 3D), for CSC-derived CS formation and their cardiomyocyte commitment in vitro. The switch from a 2D to a 3D culture microenvironment per se guides cell plasticity and myogenic differentiation within CS and is necessary for robust cardiomyocyte differentiation. On the contrary, 2D monolayer CSC cultures show a significant reduced cardiomyocyte differentiation potential compared to 3D CS culture. Forced aggregation into spheroids using hanging drop improves CS myogenic differentiation when compared to ultra-low attachment plates. Performing CS formation and myogenic differentiation exclusively in 3D culture using agarose micro-molds maximizes the cardiomyocyte yield. A 3D culture system instructs CS myogenic differentiation, thus representing a valid model that can be used to study adult cardiac regenerative biology.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 These authors contributed equally to this work. These authors also contributed equally to this work.
ISSN:	2073-4409 2073-4409
DOI:	10.3390/cells12131793