Fig. 6

Influence of VCAM-1 blockade in 2D cultured MSCs on cell aggregation, ventricular dilation and neurological function. A Cell suspensions of 2D MSCs were incubated with an anti-VCAM-1 antibody or isotype IgG and 3D MSCs treated with isotype IgG served as a control. Cell suspensions were gently loaded on a cell strainer with 40 μm pores. Cells passing through the strainer were counted, and cells retained by the filter were stained with DAPI and visualized under a fluorescence microscope. Scale bar, 100 μm (A). The results showed considerably more 3D MSCs passing through the filter than 2D MSCs, and treatment of 2D MSCs with anti-VCAM-1 antibody increased the number of cells passing through the filter by over 60% (Fig. 5B). More 2D MSC aggregates were detected on the filter than 3D MSCs, and treatment of 2D MSCs with anti-VCAM-1 antibody reduced the number of aggregates on the filter (A, C; n = 6; *P < 0.05). D–G 2D MSCs incubated with anti-VCAM-1 antibody or isotype control were intrathecally injected. Rats injected with ASCF or 3D MSCs served as controls. After 24 h, MRI analysis was performed (E; n = 6; representative images of one rat from each group are shown). The bar plot shows a decrease in the enlarged ventricle in rats receiving 2D MSCs with anti-VCAM-1 antibody (D; n = 6; *P < 0.05). The number of paw slips (F) and the average time taken to cross the beam (G) were also determined by the beam walking test (n = 6; *P < 0.05)