Connexin 37‐mediated growth regulation in primary endothelial cells (LB680)

Abstract

We have previously shown that connexin 37 (Cx37) is growth suppressive in vivo, as suggested by increased in ischemia‐induced angiogenesis and collateral vasculogenesis in Cx37‐deficient mice (Am.J.Physiol. 301: H1872‐H1881, 2011). To provide an in vitro system for studies of microvascular capillary growth control mediated by Cx37, we isolated primary skeletal muscle endothelial cells (ECs) from the hindlimb of both wild type (WT) and Cx37‐/‐ mice. Using VE‐cadherin as the positive selection marker and a live cell sorting strategy, we are able to culture primary ECs for 14‐21 days before they lose endothelial character. Evidence for endothelial nature was provided by cell morphology and the continued presence of VE‐cadherin. The growth of ECs was monitored and revealed that proliferation of ECs isolated from WT mice is slowed compared to ECs originating from Cx37‐/‐ mice. Additionally, there is a 4‐day difference between the time it takes to reach confluence of Cx37‐/‐ EC cultures and WT ECs. These data implicate Cx37 in the growth regulation of primary ECs and suggest Cx37 is growth suppressive in microvascular ECs. Future studies will provide further insight into the growth suppression mechanism by Cx37 in primary ECs.Grant Funding Source: Supported by R01HL058732 and T32HL007249