Bone marrow stromal cells (BMSCs) serve as a valuable reservoir of multipotent stem cells important in the regulation of bone homeostasis and energy metabolism. Here, we present a protocol for isolating human BMSCs (hBMSCs) and characterizing their cellular metabolism related to hBMSC functional properties. We describe steps for bioenergetics, cell senescence, and production of reactive oxygen species (ROS), together with description of the data analysis. These assays provide information on hBMSC metabolic status valuable to regenerative medicine and therapeutic applications. For complete details on the use and execution of this protocol, please refer to Tencerova et al.1.

Department of Endocrinology and Metabolism Molecular Endocrinology and Stem Cell Research Unit Odense University Hospital University of Southern Denmark Odense Denmark

Laboratory of Molecular Physiology of Bone Institute of Physiology of the Czech Academy of Sciences Prague Czech Republic

Laboratory of Molecular Physiology of Bone Institute of Physiology of the Czech Academy of Sciences Prague Czech Republic; Faculty of Science Charles University Prague Czech Republic

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