hESC Expansion and Stemness Are Independent of Connexin Forty-Three-Mediated Intercellular Communication between hESCs and hASC Feeder Cells
Figure 2
Functional gap junction channels were detected by the scrape loading/dye transfer assay for hASCs cultured alone and with hESCs. Phase contrast (A(a-c), B(a-c), C(a,b)), Lucifer yellow (A(d-f), B(d-f), C(c,d)) and rhodamine-dextran (A(g-i), B(g-i), C(e,f)) of hASCs and hESCs are shown. A scrape was made across the monolayer to allow the uptake of Lucifer yellow by the wounded cells. The dye was then transferred from the wounded cells to the adjacent, intact cells via functional gap junction channels. Lucifer yellow dye transfer was shown from hASCs to adjacent hASCs (A(e,f)); hESCs to adjacent hESCs (B(e,f); and hASCs to adjacent hESCs (C(c,d)). No-scrape group was employed as the normal conditions (A(d) and B(d)). Rhodamine-dextran was used as a negative control, showing no dye transfers from the wounded cells to neighbouring cells (A(h,i), B(h,i), C(e,f)). The black and white arrows indicate the scraped cells and the hESC colonies, respectively. The white dotted lines show the boundary between the hASCs and hESC colony. Scale bar, 100 μm.