Figure 2

HSCs promote chemoresistance and compactness in multicellular tumor spheroids (MCTS).

(A,B) Drug sensitivity was measured by EthD-1 (dead cell marker) staining in HepG2 spheroids (A) and Huh7 spheroids (B), which were co-cultured with or without LX2, WI38, and HUVECs and treated with 10 μM sorafenib for 4 days (upper panel). The intensity of EthD-1 was analyzed and values were normalized to control (0.01% DMSO) (lower panel). Data are shown as means ± SD from three independent experiments with duplicates. (C) HepG2 spheroids with or without LX2, WI38, and HUVECs at a 1:1 ratio were treated with 10 μM cisplatin for 4 days (upper panel). Intensity of EthD-1 was examined and values were normalized to control (0.01% DMSO) (lower panel). Data are shown as means ± SD from three independent experiments with duplicates. (D) HepG2 spheroids alone or with LX2 or WI38 were treated with 10 μM doxorubicin for the indicated number of hr. The penetration of doxorubicin into the spheroids was measured with LSM with various confocal stacks. (E) 10 μM of doxorubicin was treated in HepG2 spheroids alone or with LX2 or WI38 for indicated time. Absorbance of doxorubicin, which was penetrated in each spheroid, was examined. Values were normalized to 1 hr. Data are shown as means ± SD from two independent experiment with duplicate. *P < 0.05, **P < 0.005. Scale bar = 200 μm.