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3D （Three-Dimensional） Caco-2 Spheroids： Optimized in vitro protocols.to Favor Their Differentiation Process and to Analyze Their Cell Growth Behavior

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Journal of Pharmacy and Pharmacology 2016年第7期4卷 341-350页

作者： Gabriella Rainaldi Alessandra Boe Sandra Gessani Department of Hematology Oncology and Molecular Medicine lstituto Superiore di Sanit& Rome Italy

3D （Three-dimensional） Caco-2 spheroids closely recapitulating in vivo physiological organization of intestinal epithelial cells, provide an excellent in vitro model system to study their pathophysiology and their response to stressful stimuli. The objective of this technical note is to provide optimized in vitro experimental protocols.for culturing 3D Caco-2 spheroids and for analyzing their cell growth features. An optimized 3D Caco-2 spheroid culturing technique based on a new configuration of the culture medium is provided A methodological approach to determine the distribution of the cell cycle phases in disaggregated Caco-2 spheroids by using cytofluorimetric analysis is also described. The optimized culturing protocol favors 3D Caco-2 spheroid differentiation process, as evaluated by the number of well-differentiated spheroids with a single hollow lumen. The cytofluorimetric analysis allows rapid collection of cell cycle phase data from high numbers of spheroid samples, thus, permitting to estimate their growth dynamics in a relatively short time. The optimized technical approaches described here can be applied in systematic manner to a variety of research activities utilizing 3D Caco-2 spheroids. Ease of use, time and economic saving advantages deriving from these protocols.further highlight their potential.

关键词： 3D multicellular spheroids 3D intestinal epithelial spheroids 3D Caco-2 spheroid culture systems optimizedexperimental protocols.

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