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Mouse brown adipose tissue was dissociated using the Adipose Tissue Dissociation Kit, mouse and rat (# 130-105-505) in combination with the gentleMACS™ Octo Dissociator with Heaters. Subsequently, brown adipose tissue derived progenitor cells were isolated using the Adipocyte Progenitor Isolation Kit, mouse. Cells were fluorescently stained with lineage markers (CD31/CD45/Terr119), and Anti-Sca-1-FITC and analyzed using the MACSQuant ® Analyzer. |
Unseparated fraction | Enriched adipocyte progenitor cells |
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Isolated progenitor cells were cultured in expansion medium (DMEM, 10% FCS, 10 ng/mL human FGF-2, 100 U/mL penicillin/ 100 U/mL streptomycin) for four days. After four days, medium was replaced with differentiation medium (DMEM, 4 nM Insulin, 1 µM Rosiglitazone, 10%FCS, 100 U/mL penicillin/ 100 U/mL streptomycin) and cells were incubated for three more days to induce differentiation into adipocytes. Cells were stained with Nile Red to identify lipid storage. |
Phase contrast image of adipocytes | Nile Red stained differentiated adipocytes |
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