Isolation of Murine Adipose-Derived Stromal/Stem Cells for Adipogenic and Osteogenic Differentiation or Flow Cytometry-Based Analysis

Abstract

Murine models of obesity or reduced adiposity are a valuable resource for understanding the role of adipocyte dysfunction in metabolic disorders. Adipose tissue stromal vascular cells or primary adipocytes derived from murine adipose tissue and grown in culture are essential tools for studying the mechanisms underlying adipocyte development and function. Herein, we describe methods for the isolation, expansion, and long-term storage of murine adipose-derived stromal/stem cells, along with protocols for inducing adipogenesis to white or beige adipocytes in this cell population and osteogenic differentiation. Isolation of the adipose stromal vascular fraction cells for flow cytometric analysis is also described.

Keywords: Adipogenesis; Adipose-derived stromal/stem cells (ASCs); Beige adipogenesis; Collagenase; Flow cytometry; Hematopoietic stem cells (HSCs); Isolation; Mesenchymal stem cells (MSCs); Murine; Osteogenesis; Stromal vascular fraction (SVF).