The Adipocyte Progenitor Isolation Kit, mouse has been designed for the isolation of tissue-derived progenitor cells from dissociated brown and white adipose tissue. For optimal results, the Adipocyte Progenitor Isolation Kit, mouse should be used in combination with the Adipose Tissue Dissociation Kit, mouse and rat (# 130‑105‑808).
In mammals the two main types of adipose tissues exist, white adipose tissue (WAT) and brown adipose tissue (BAT). Both types fulfil different physiological functions. White adipose tissue (WAT) is for storing energy, while brown adipose tissue (BAT) is for energy consumption. Using the Adipose Tissue Progenitor Isolation Kit, mouse adipose tissue derived progenitor cells are isolated by depletion of non-target cells followed by positive selection of the target cells. Non-target cells are directly magnetically labeled with a cocktail of monoclonal antibodies conjugated with MACS MicroBeads. The magnetically labeled non-target cells are depleted by retaining them within a MACS Column in the magnetic field of a MACS Separator, while the unlabeled adipose tissue progenitor cells pass through the column. Subsequently, the progenitor cells are directly magnetically labeled. The magnetically labeled target cells are retained within a MACS Column in the magnetic field of a MACS Separator, while the unlabeled non-target cells pass through the column.
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.
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