Human Amniotic Fluid Cells Form Functional Gap Junctions with Cortical Cells

Figure 6

AF cells repair the scratch-induced wound injury in cortical cultures. (a)-(b) A low magnification of confluent cortical cultures stained with Hoechst (a and b, blue), GFAP (b, red) and Cx43 (b, green). (c)-(d) Parallel cortical cultures were subjected to scratch injury and stained with the same markers 24 hrs later. GFAP positive astrocytes were seen at the scratch border (d), without repairing the wound. (e)-(f) In contrast, when AF cells were seeded following scratch, they filled the injury site (f, arrows), maintained CX43 expression (f, green), and facilitated wound repair. (g)–(k) To further identify AF cells located in the injury site, separate cultures were stained with human mitochondrial marker (red), CX43 (green), and Hoechst counter stain (blue). Cx43 was readily expressed at the boundary between AF cells and cortical cells (k). (l)–(n) Live assays, using GFP-tagged AF cells, were also used to confirm wound repair after scratch injury in cortical cultures. Scale bar: 150 μm (a)–(f), 35 μm (g)-(h), 90 μm (I, J), 10 μm (k), 80 μm (l)–(n).