Effects of exogenous sonic hedgehog peptide on proliferation, adhesion, migration of endothelial progenitor cells from peripheral blood.

In order to study the effects of exogenous sonic hedgehog (shh) peptide on proliferation, adhesion, migration of endothelial progenitor cells (EPCs) from rat peripheral blood, the mononuclear cells were collected from rat peripheral blood by Ficoll density gradient centrifugation. EPCs were isolated with adherence screening method and cultured in M199 culture medium with the supplement of VEGF and bFGF. The immunohistochemical staining was used to identify cell markers such as CDl33 and VEGFR-2. EPCs were stimulated with exogenous shh peptide of different final concentrations (0.01, 0.1, 1, 10 μg/mL). The proliferation, adhesion and migration of EPCs were detected by MTT chromometry, adhesion test and transwell system, respectively. The results of this study showed that, after 7 days of culture, cells formed clusters, assuming typical cobbles-tone pattern under microscope. After 2 weeks of culture, cells were arranged in cord-like fashion and sometimes grew like "micro-vessels". Immunohistochemical staining showed that the cultured cells were positive for both CD133 and VEGFR-2. The proliferation, adhesion and migration of EPCs could be promoted by endogenous shh peptide at concentrations from 0.1 μg/mL to 10 μg/mL in a concentration-dependent manner. The findings indicate that exogenous shh peptide can enhance EPCs proliferation, adhesion, and migration, which may have a potential value for clinical application.