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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： Caco-2细胞肠上皮细胞优化培养分化过程体外培养球体协议三维

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Dietary fatty acids and adipose tissue inflammation at the crossroad between obesity and colorectal cancer

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Journal of Cancer Metastasis and Treatment 2019年第8期4卷 29-40页

作者： Lucia Conti Manuela Del Cornò Beatrice Scazzocchio Rosaria Varì Massimo D'Archivio Barbara Varano Roberta Masella Sandra Gessani Center for Gender-specific Medicine Istituto Superiore di Sanità 00161 Rome Italy

Excess adiposity, a worldwide-growing pathological condition, is now recognized as a main risk factor for most chronic diseases including colorectal cancer (CRC). Obese subjects show an increased cancer incidence with obesity representing an important indicator of survival, prognosis, recurrence and response to therapy. A low-grade chronic inflammation of metabolically active tissues including the adipose tissue (AT), defined as meta-inflammation, is a main feature of obesity. Fatty acids (FA), the main AT components, are important modulators of inflammation, and the type of FA stored in AT critically affects tissue functions. Their profile within AT mirrors FA dietary intake but also depends on a metabolic control. Obesity, changes in the habitual diet, weight loss or pathological conditions like CRC influence FA profile of AT pointing to these molecules as important actors in AT dysfunction and meta-inflammation, that characterize metabolic diseases and may favor cancer development. Worth of note, diet is receiving growing attention as a main determinant in cancer prevention due to its capacity to modulate immune response and inflammation. Alterations in the balance between different families of FA may contribute to generate a pro-inflammatory profile with deleterious effects on metabolic and immune homeostasis at both local and systemic levels. This review focuses on FA as regulators of human AT inflammation discussing the role of obesity-, diet-, and weight loss-associated changes in FA profile in this process. The relevance of FA composition of AT in linking diet, obesity and CRC will be also reviewed.

关键词： Obesity adipose tissue inflammation diet weight loss fatty acids colorectal cancer

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