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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®
Enriched adipocyte progenitor cells
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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