Integration of endothelial cells in multicellular spheroids prevents apoptosis and induces differentiation
Thomas Korff,Hellmut G. Augustin +1 more
TLDR
The findings indicate that polarized surface EC differentiate to become independent of exogenous survival factors and demonstrate that spheroid cell culture systems are useful not just for the study of tumor cells and embryonic stem cells but also for the analysis of differentiated functions of nontransformed cells.Abstract:
Single endothelial cells (EC) seeded in suspension culture rapidly undergo apoptosis. Addition of survival factors, such as VEGF and FGF-2, does not prevent apoptosis of suspended EC. However, when cells are allowed to establish cell–cell contacts, they become responsive to the activities of survival factors. These observations have led to the development of a three-dimensional spheroid model of EC differentiation. EC spheroids remodel over time to establish a differentiated surface layer of EC and a center of unorganized EC that subsequently undergo apoptosis. Surface EC become quiescent, establish firm cell–cell contacts, and can be induced to express differentiation antigens (e.g., induction of CD34 expression by VEGF). In contrast, the unorganized center spheroid cells undergo apoptosis if they are not rescued by survival factors. The responsiveness to the survival factor activities of VEGF and FGF-2 was not dependent on cell shape changes since it was retained after cytochalasin D treatment. Taken together, these findings characterize survival factor requirements of unorganized EC and indicate that polarized surface EC differentiate to become independent of exogenous survival factors. Furthermore, they demonstrate that spheroid cell culture systems are useful not just for the study of tumor cells and embryonic stem cells but also for the analysis of differentiated functions of nontransformed cells.read more
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Isolation and Expansion of Adult Cardiac Stem Cells From Human and Murine Heart
Elisa Messina,Luciana De Angelis,Giacomo Frati,Stefania Morrone,Stefano Chimenti,Fabio Fiordaliso,Monica Salio,Massimo Battaglia,Michael V.G. Latronico,M. Coletta,E. Vivarelli,Luigi Frati,Giulio Cossu,Alessandro Giacomello +13 more
TL;DR: The isolation of undifferentiated cells that grow as self-adherent clusters (that are termed “cardiospheres”) from subcultures of postnatal atrial or ventricular human biopsy specimens and from murine hearts are described.
Journal ArticleDOI
Spheroid-based drug screen: considerations and practical approach
TL;DR: This work describes a standardized setup for reproducible, easy-handling culture, treatment and routine analysis of multicellular spheroids, the classical 3D culture system resembling many aspects of the pathophysiological situation in human tumor tissue and provides a list of human carcinoma cell lines that produce treatable sp Heroids under identical culture conditions.
Journal ArticleDOI
Recent advances in three-dimensional multicellular spheroid culture for biomedical research
Ruei-Zhen Lin,Hwan-You Chang +1 more
TL;DR: The current understanding of multicellular spheroid formation mechanisms, their biomedical applications, and recent advances in sp heroid culture, manipulation, and analysis techniques are reviewed.
Journal ArticleDOI
Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy.
TL;DR: The suitability of spheroids as an in vitro platform for testing drug delivery systems is examined and the assay techniques required for the characterization of drug delivery and efficacy in sp Heroids are discussed.
Journal ArticleDOI
Role of Dicer and Drosha for endothelial microRNA expression and angiogenesis.
TL;DR: In this paper, the role of Dicer and Drosha for angiogenesis was investigated and it was shown that genetic silencing of both enzymes significantly reduced capillary sprouting of endothelial cells and tube forming activity.
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