Fig. 3. Cellular uptake of non-incubated EVs and HC-EVs in phagocytic and HepG2 cells.
a–f, Cellular uptake of EV2D ± HC (a–c) and EV3D ± HC (d–f) in phagocytic J774 cells (a,d), human-monocyte-derived macrophages (Hu-Ø) (b,e) and HepG2, representing non-phagocytic liver hepatocytes (c,f). AF488-labelled EVs or HC-EVs were incubated with cells at a dose of 2 × 109 particles per well (24-well plate) for 1 h, 4 h and 24 h. Cellular uptake was measured by flow cytometry, and uptake was expressed as fold increase of the mean AF488 signal per cell (MFI) compared with non-treated cells. Doping of EVs with albumin-rich HC, that is, the case of EV3D, significantly reduces uptake in phagocytic cells while keeping uptake in HepG2 cells unchanged (1**P = 0.00417, 2***P = 0.00021, 3**P = 0.00243, 4*P = 0.04176, 5**P = 0.00871, 6****P = 0.00003, 7**P = 0.00522). Data are presented as mean ± s.d. (n = 3) with two-tailed unpaired t-test (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001).