Figure 4

hMSC migrate to the AEC wound sites and close wound gaps. (A) Schematic diagram showing two-cell direct contact co-culture wound repair model using 3 μm porous transwell system. (B) Laser scanning confocal micrographs of AEC-MSC direct contact co-culture wound repair model at 0 and 24 hours. Red cells are DiI labelled AECs and green cells are DiO labelled hMSCs. Horizontal panels are the Z-slicing through the wound gaps and the vertical panels are Z-slicing through the corresponding juxta-wound monolayers. Appearance of green signals in the same plane of red signals in the in the Z-slice after 24 hours confirms hMSC migration to the AEC wound site. (C) AEC-MSC direct contact co-culture vs. negative and positive controls. Black and grey bars indicate AEC and hMSC contribution to repair, respectively. Negative control represents AEC wound repair in SF-DMEM in mono-culture setting on upper surface of transwell PET membrane. Positive control represents AEC wound repair in 10% FBS supplemented DMEM in mono-culture setting on upper surface of transwell PET membrane. (n = 6; ***p < 0.001). (D) hMSC migration in a direct contact co-culture wound repair system (n = 13; ***p < 0.001). Data presented as mean ± SD. Scale bar, 150 μm.