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A simple method to sort ESC‐derived adipocytes
Author(s) -
Schaedlich Kristina,
Knelangen Julia M.,
Navarrete Santos Alexander,
Fischer Bernd,
Navarrete Santos Anne
Publication year - 2010
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20953
Subject(s) - cell sorting , adipogenesis , adipocyte , adipose tissue , microbiology and biotechnology , flow cytometry , embryoid body , cell , biology , embryonic stem cell , stem cell , cellular differentiation , mass cytometry , cell culture , chemistry , biochemistry , phenotype , adult stem cell , genetics , gene
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