Fig. 4

3D MSCs exhibited enhanced anti-inflammatory effects on microglia in vivo. a The expression of stanniocalcin 1 (STC1), hepatocyte growth factor (HGF), and tumor necrosis factor-stimulated gene-6 (TSG-6) between 2D and 3D MSCs was evaluated with qRT-PCR. b Different morphology of ionized calcium-binding adapter molecule 1 (Iba1)-positive microglia with magnified images at 5 days after ischemic stroke. c Immunofluorescence imaging of microglia and glial fibrillary acidic protein (GFAP)-positive astrocytes in the ischemic core at 5 days after MCAO. The number of amoeboid Iba1-positive cells was reduced in the 3D MSC-administered group. d The quantitative analysis of amoeboid Iba1-positive cells indicated that the number of amoeboid cells in the ischemic core was significantly decreased in the 3D MSC-administered group than that in the PBS and 2D MSCs group. Data are shown as mean ± SEM; n = 3 for each group. **p < 0.01. e, f qRT-PCR analysis showed that IV injection of 3D MSCs significantly decreased the expression of Iba1, CD45, interleukin-1β (IL-1β), and interleukin-6 (IL-6) compared with the PBS and 2D MSCs group. Data are shown as mean ± SEM; n = 3 for each group. *p < 0.05, **p < 0.01, ***p < 0.001