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Efficient in vitro adipocyte model of long-term lipolysis: A tool to study the behavior of lipophilic compounds

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Abstract

The triglycerides (TGs) stored in the white adipose tissue are mobilized during periods of negative energy balance. To date, there is no in vitro model of adipocytes imitating a long period of negative energy balance in which triglycerides are highly mobilized. Such model would allow studying the mobilization of TGs and lipophilic compounds trapped within the adipose tissue (e.g., pollutants and vitamins). The present study aims at developing a performing long-term in vitro lipolysis in adipocytes, resulting in a significant decrease of TG stores. Lipolysis was induced on differentiated rat adipocytes by a lipolytic medium with or without isoproterenol for 12 h. The condition with isoproterenol was duplicated, once with medium renewal every 3 h and once without medium renewal. Adding isoproterenol efficiently triggered lipolysis in a short time (3 h). However, a single stimulation by isoproterenol, without medium renewal, was not sufficient to reduce the TG content during a longer term (12 h). A reesterification of fatty acids occurred after a few hours of lipolysis, resulting in a novel increase of cellular lipids. Regular medium renewal combined with repeated isoproterenol stimulations led to almost emptied cells after 12 h. However, medium renewal without isoproterenol stimulation for 12 h was as efficient in terms of lipid mobilization. Our study demonstrates that, over a short-term period, isoproterenol is required to exert a significant lipolytic effect on adipocytes. During a long-term period, the presence of isoproterenol is no longer essential. Instead, medium renewal becomes the main factor involved in cell emptying. The efficiency of this protocol was demonstrated by visual tracking of the cells and by monitoring the dynamics of release of a lipophilic compound, PCB-153, from adipocytes during lipolysis.

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Acknowledgments

The authors are very grateful to Marie-Thérèse Ahn, Daniel Jal, Willy Marteau, and Philippe Bombaerts from “Institut des Sciences de la Vie” (ISV), UCLouvain, for their technical assistance. We also thank Pauline Beguin, Julie Winand, and Anne-Catherine Schneider from ISV for their help in the fatty acid analyses. Members of “Support en méthodologie et calcul statistique” (Institut multidisciplinaire pour la modélisation et l’analyse quantitative, UCLouvain, Belgium) are gratefully acknowledged for the collaboration in the statistical analyses. We also greatly appreciated the help and advice of Guillaume Bernard for picture processing.

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  1. Institut des Sciences de la Vie, UCLouvain, Croix du Sud 2/L7.05.08, 1348, Louvain-la-Neuve, Belgium

    Caroline Louis, Carine Van den Daelen, Gilles Tinant, Sophie Bourez, Isabelle Donnay, Yvan Larondelle & Cathy Debier

  2. Laboratoire d’Ecologie animale et d’Ecotoxicologie, Université de Liège, Allée du 6 août 15, 4000, Liège, Belgium

    Jean-Pierre Thomé

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  1. Caroline Louis
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Louis, C., Van den Daelen, C., Tinant, G. et al. Efficient in vitro adipocyte model of long-term lipolysis: A tool to study the behavior of lipophilic compounds. In Vitro Cell.Dev.Biol.-Animal 50, 507–518 (2014). https://doi.org/10.1007/s11626-014-9733-6

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