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Flow Cytometric Isolation and Differentiation of Adipogenic Progenitor Cells into Brown and Brite/Beige Adipocytes

  • Jochen Steinbring
  • Antonia Graja
  • Anne-Marie Jank
  • Tim J. Schulz
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1566)

Abstract

Aside from mature adipocytes, adipose tissue harbors several distinct cell populations including immune cells, endothelial cells, and adipogenic progenitor cells (AdPCs). AdPCs represent the reservoir of regenerative cells that replenishes adipocytes during normal cellular turnover and during times of increased demand for triglyceride-storage capacity. The worldwide increase in pathologies associated with the metabolic syndrome, such as obesity and type-2 diabetes, has heightened public and scientific interest in adipose tissues and the cell biological processes of adipose tissue formation and function. Two distinct types of fat cells are known: White and brown adipocytes. Especially brown adipose tissue (BAT) has received considerable attention due to its unique capacity for thermogenic energy expenditure and potential role in the treatment of adiposity. Accordingly, the cold-induced conversion of white into brown-like adipocytes has become a feasible approach in humans and a study-subject in rodents to better understand the underlying molecular processes. Fluorescence-activated cell sorting (FACS) provides a method to isolate AdPCs and other cell populations from adipose tissue by using antibodies detecting unique surface markers. We here describe an approach to isolate cells committed to the adipogenic lineage and summarize established protocols to differentiate FACS-purified primary AdPCs into UCP1-expressing brown adipocytes under in vitro conditions.

Key words

Brown/brite adipose tissue White adipose tissue Adipogenic progenitor cells Fluorescence-activated cell sorting Cell surface marker antibodies Brown adipogenesis 

Notes

Acknowledgments

This work was supported by grants from the German Research Foundation (DFG; grant # SCHU 2445/2-1) and the European Research Council (grant # ERC-StG-2012-311082) to T.J.S. The authors gratefully acknowledge support from the German Center for Diabetes Research (DZD).

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Jochen Steinbring
    • 1
  • Antonia Graja
    • 1
  • Anne-Marie Jank
    • 1
  • Tim J. Schulz
    • 1
    • 2
  1. 1.Research Group Adipocyte DevelopmentGerman Institute of Human Nutrition (DlfE)NuthetalGermany
  2. 2.German Center for Diabetes Research (DZD)München-NeuherbergGermany

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