Exosome-Mediated BMSC Proliferation

Cell proliferation was detected using the CCK-8 (Solarbio, Beijing, China, CA1210) assay. According to the manufacturer’s instructions, we made BMSCs cell suspensions of 5×10⁴/mL, injected into 96-well plate, cultured in 5% CO₂ and 95% O₂ in the incubator at 37° C. Divided into four treatment groups: (1) Control group; (2) PMO group; (3) PMO+Exos group; and (4) Exos Group, with three replicates in each group. The quantity of exosome added to BMSCs for the treatments was consistent with previous reports [19 (link)]. Control Group and PMO Group were added with Exo-free a-MEM medium (Gibco, NY, USA, C12571500BT), PMO+Exos Group (PMO BMCs treated with exosomes) and Exos Group (normal BMCs treated with exosomes) were added with 100μg/ mL exosome a-MEM medium (Gibco, NY, USA, C12571500BT). All groups were replaced with new a-MEM medium (Gibco, NY, USA, C12571500BT) as them required. The cell plate was cultured in the incubator, and 10 μL of CCK-8 (Solarbio, Beijing, China, CA1210) solution was added into each well and incubated in the incubator for 4 hours at 0 h, 24 h, 48 h and 72 h. An enzyme standard instrument (ThermoFisher Scientific, NY, USA, Multiskan SkyHigh) was used to measure the absorbance at 450 nm.