Supplementary material from "Cytoplasmic flow induced by a rotating wire in living cells: Magnetic rotational spectroscopy and finite element simulations "

Recent studies have highlighted intracellular viscosity as a key biomechanical property with potential as a biomarker for cancer cell metastasis. In the context of cellular mechanobiology, magnetic rotational spectroscopy (MRS), which employs rotating magnetic wires of length L = 2-8 µm to probe cytoplasmic rheology, has emerged as an effective method for quantifying intracellular viscoelasticity. This study examines microrheology data from three breast epithelial cell lines, MCF-10A, MCF-7, and MDA-MB-231, along with new data from HeLa cervical cancer cells. Here, MRS is combined with finite element simulations to characterize the flow field induced by wire rotation in the cytoplasm. COMSOL simulations performed at low Reynolds numbers show that the flow velocity is localized around the wire, and display characteristic dumbbell-shaped profiles. For wires representative of MRS experiments in cells, the product of shear rate and cytoplasmic relaxation time (γ̇τ with τ ~ 1 s) remains below unity, indicating that the flow occurs within the linear regime. This outcome confirms that MRS can reliably measure the zero-shear viscosity of the intracellular medium in living cells. This study also demonstrates that integrating MRS intracellular measurements with COMSOL simulations significantly improves the reliability of in vitro assessments of cytoplasmic mechanical properties.