Human Amniotic Fluid Cells Form Functional Gap Junctions with Cortical Cells

Figure 2

Connexin expression in c-kit-positive and single-cell-derived AF clones. (A) c-kit-positive AF cells were obtained by fluorescence activated cell sorting (FACS). (B) RT-PCR analysis of connexin (CX) expression in c-kit-positive (AF-c-kit) and single-cell-derived (AF-F5) AF clones. GAPDH transcript was used as an internal control. (C) Immunocytochemistry confirmed the expression of CX43 (green) in AF-c-kit and AF-F5 cultures. Hoechst was used as a counter-stain (blue). (b) and (d) represent the corresponding phase contrast images. Scale bar: 25 μm. (D) AF donor cells were preloaded with DiI (red) and calcein (green) and plated as single cells on cultures of AF cells. Calcein transferred from the donor AF cell to neighbouring cells, confirming the formation of functional gap junctions among AF-c-kit cells as well as AF-F5 cells within 4 hours. Scale bar: 50 μm.