A simple method to sort ESC-derived adipocytes

Authors

  • Kristina Schaedlich,

    1. Department of Anatomy and Cell Biology, Martin Luther University, Faculty of Medicine, Halle, Germany
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    • Kristina Schaedlich and Julia M. Knelangen contributed equally to this work.

  • Julia M. Knelangen,

    1. Department of Anatomy and Cell Biology, Martin Luther University, Faculty of Medicine, Halle, Germany
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    • Kristina Schaedlich and Julia M. Knelangen contributed equally to this work.

  • Alexander Navarrete Santos,

    1. Department of Cardiothoracic Surgery, Martin Luther University, Faculty of Medicine, Halle, Germany
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  • Bernd Fischer,

    1. Department of Anatomy and Cell Biology, Martin Luther University, Faculty of Medicine, Halle, Germany
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  • Anne Navarrete Santos

    Corresponding author
    1. Department of Anatomy and Cell Biology, Martin Luther University, Faculty of Medicine, Halle, Germany
    • Martin Luther University, Faculty of Medicine, Department of Anatomy and Cell Biology, Grosse Steinstrasse 52, D-06097 Halle (Saale), Germany
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Abstract

Because of the increasing incidence of worldwide obesity, cell culture models which enable the study of adipose tissue development are of particular importance. The murine embryonic stem cell (ESC) line CGR8 differentiates into adipocytes with a differentiation efficiency of up to 15%. A critical step for the analysis of stem cell-derived adipogenesis is the reliable separation of adipocytes. Here we report on how to (i) gently separate the cells of embryoid bodies (EBs) and (ii) identify and sort adipocytes from the rest of the heterogeneous cell mixture. Up to the present, no adipocyte specific surface marker is known for fluorescence activated cell sorting (FACS). After separation we employed two independently existing FACS methods for adipocyte cell sorting. These methods are based on Nile red staining and granularity. For stem cell-derived adipocytes only the combination of both methods led to a reliable, efficient, and highly reproducible FACS analysis, as shown by the presence and absence of adipocyte specific markers in positively and negatively sorted cells. © 2010 International Society for Advancement of Cytometry

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