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Techniques and Methods

Ex vivo microperfusion system of the adipose organ: a new approach to studying the mobilization of adipose cell populations

International Journal of Obesity volume 38, pages 1255–1262 (2014)Cite this article

Subjects

Abstract

Backround/Objectives:

Adipose tissue (AT) is a dynamic organ that expands and contracts rapidly. It is composed of adipocytes and of cell populations among which immune cells and mesenchymal progenitors known as adipose stromal cells (ASCs). The AT cell turnover has been extensively studied. Surprisingly it has only been viewed as the result of both cell proliferation/death and cell infiltration. Nevertheless, both immune cells and ASCs exhibit migration abilities; therefore their egress from AT in response to physiological/pathophysiological stimuli has to be considered. To do so, the aim of the present work was to develop a model allowing the study of cell release from the adipose organ.

Subjects/Methods:

Mesenteric (Mes) ATs were isolated from 9-week-old C57BL/6 male mice and were catheterized via the superior mesenteric artery and were perfused with a saline solution. After an equilibration period, the mesenteric fat pad was perfused with CXCL12 (stromal-derived factor-1, SDF-1) or sphingosine 1-phosphate (S1P) to trigger cell mobilization and perfusates were collected every 30 min for subsequent flow cytometry analyses.

Results:

We report here that CXCL12 induces the specific release of ASCs from MesAT thus demonstrating that ASCs are specifically mobilized from fat depots by a CXCL12-dependent pathway. Moreover, we showed that leukocyte mobilization can be triggered via a S1P-dependent pathway.

Conclusions:

We have developed a microperfusion model of an intact fat depot allowing the study of AT cell release in response to various molecules. The perfusion system described here demonstrates that ASCs and leukocytes can be pharmacologically mobilized from AT. Therefore, AT microperfusion might constitute an appropriate and reliable approach for evaluating the mobilization of different cell populations from AT in various physiological and pathophysiological contexts. Such a model might help in identifying factors and drugs controlling AT cell release, impacting the medical fields of regenerative medicine and of obesity or its associated comorbidities.

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Acknowledgements

This work was supported by Inserm, ‘La ligue nationale contre le cancer’, the Association Française d’étude et de recherches sur l’obésité (Afero) and Roche (2007). We would like to thank Christophe Guissard for his invaluable technical assistance. We are grateful to the ‘Institut des Technologies Avancées en sciences du Vivant’ (ITAV) UMS 3039 and Dr J Rouquette for adipose tissue imaging.

Author information

Authors and Affiliations

  1. Inserm U1031 STROMAlab BP 84 225-F-31 432, Toulouse, France

    M Gil-Ortega, L Casteilla & C Sengenès

  2. CNRS, Université Toulouse III, UPS UMR5273 STROMAlab, BP 84 225-F-31 432, Toulouse, France

    M Gil-Ortega, L Casteilla & C Sengenès

  3. EFS (Etablissement Français du Sang), STROMAlab BP 84 225-F-31 432, Toulouse, France

    M Gil-Ortega, L Casteilla & C Sengenès

  4. Université Toulouse III, UPS UMR5273 STROMAlab BP 84 225-F-31 432, Toulouse, France

    M Gil-Ortega, L Casteilla & C Sengenès

  5. Instituto Pluridisciplinar, Facultad de Farmacia, Universidad Complutense de Madrid, Juan XXIII 1, Madrid, 28040, Spain

    M S Fernández-Alfonso

  6. Departamento de Ciencias Farmacéuticas y de la Salud, Facultad de Farmacia, Universidad CEU-San Pablo, Madrid, 28668, Spain

    B Somoza

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Correspondence to M Gil-Ortega.

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Gil-Ortega, M., Fernández-Alfonso, M., Somoza, B. et al. Ex vivo microperfusion system of the adipose organ: a new approach to studying the mobilization of adipose cell populations. Int J Obes 38, 1255–1262 (2014). https://doi.org/10.1038/ijo.2013.243

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