OBJECTIVE: To study the effect of shh on the migration, proliferation and phenotypic modulation of vascular adventitial fibroblasts. METHOD: Cultivate the vascular adventitial fibroblast in vitro. Use immunofluorescent, laser confocal microscopy, Western-blot and real-time PCR to detect the expression of mRNA and protein of related index. Estimate cell proliferation according to the expression of Ki67 and cell proliferation curve. Application of wound healing test to estimate migration of fibroblast. The expression of alpha-actin is thought to be marker of phenotypic modulation of fibroblast. RESULT: The expression of shh was detected in vascular adventitial fibroblast in vitro. After addition of exogenous shh (3.5 microg/ml), there were more Ki67(+) cells and the wounding area which was covered by cells became larger. The expression of alpha-actin was detected. After addition of cyclopamine (40 micromol/L), there were less Ki67(+) cells and the wounding area which was covered by cells became smaller. CONCLUSION: Shh can promote proliferation, migration and phenotypic modulation of vascular adventitial fibroblasts.
OBJECTIVE: To study the effect of shh on the migration, proliferation and phenotypic modulation of vascular adventitial fibroblasts. METHOD: Cultivate the vascular adventitial fibroblast in vitro. Use immunofluorescent, laser confocal microscopy, Western-blot and real-time PCR to detect the expression of mRNA and protein of related index. Estimate cell proliferation according to the expression of Ki67 and cell proliferation curve. Application of wound healing test to estimate migration of fibroblast. The expression of alpha-actin is thought to be marker of phenotypic modulation of fibroblast. RESULT: The expression of shh was detected in vascular adventitial fibroblast in vitro. After addition of exogenous shh (3.5 microg/ml), there were more Ki67(+) cells and the wounding area which was covered by cells became larger. The expression of alpha-actin was detected. After addition of cyclopamine (40 micromol/L), there were less Ki67(+) cells and the wounding area which was covered by cells became smaller. CONCLUSION:Shh can promote proliferation, migration and phenotypic modulation of vascular adventitial fibroblasts.
Authors: Ciaran J Mooney; Roya Hakimjavadi; Emma Fitzpatrick; Eimear Kennedy; Dermot Walls; David Morrow; Eileen M Redmond; Paul A Cahill Journal: Stem Cells Int Date: 2015-05-06 Impact factor: 5.443