Abstract: Stem cells (SCs) are a class of self-renewing cells capable of differentiating into various human cell types. The field of stem cell regenerative medicine is flourishing, with China's first approved stem cell drug—Amimestrocel—bringing new hope to patients. Current research on stem cell mechanisms primarily focuses on biological and chemical factors; however, physical regulatory factors (e.g., mechanical forces, electromagnetic fields) offer distinct advantages, including non-invasiveness and temporally precise control. This review explores mechanisms by which physical factors regulate stem cell behavior. Dynamic mechanical forces influence stem cell differentiation and function by modulating key signaling pathways such as TGFβ/Smad, RAS/MAPK, and JAK/STAT, the activation of which plays crucial roles in organismal development and disease pathogenesis. Electromagnetic fields regulate stem cell proliferation, migration, and extracellular matrix remodeling through the modulation of membrane potential, ion channel activity, and gene expression, opening new research directions. Photobiomodulation enables stem cell regulation through adjustments to light parameters (wavelength, intensity, duration) or photosensitive materials, demonstrating significant effects in studies involving neural stem cells and bone marrow mesenchymal stem cells. Synergistic therapeutic strategies combining multiple physical factors with emerging technologies such as 3D printing and nanomaterials will further advance stem cell research and clinical applications.