Direct communication between human umbilical vascular endothelial cells cultured in vitro

Objective To find the morphological proof of direct communication between human umbilical vascular endothelial cells (HUVECs). Methods mRNA and protein of connexin Cx43 in cultured HUVECs were detected by RT-PCR and immunofluorescence staining. Scrape-loading dye transfer method was used to display the direct communication between HUVECs. Results Cx43 mRNA was approved to express in cultured HUVECs by RT-PCR. Protein of Cx43, located mainly on the membrane of HUVECs, but a little in the cell plasma, was also revealed by immunofluorescence staining. In the Scrape-loading dye transfer experiment, a small molecular dye, Lucifer Yellow CH, was observed to migrate between HUVECs. However, the large molecular dye of Rhodamine could not migrate to any other none loaded HUVECs. Lucifer Yellow CH could not freely transverse cell membrane except via gap junction direct communication. Conclusion The results have provided the new morphological proof for direct communication between cultured HUVECs.