Purchase this article with an account.
Kyu Yeon Han, Jin-Hong Chang, Dimitri T. Azar; Characterization of Vegf-c Induced Lymphatic Endothelial Cell Signaling By Bioelectrical Impedance Analysis. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4827.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To calculate impedance changes in human lymphatic endothelial cells after stimulation with VEGF-C in the presence and absence of MAP kinase inhibitors.
Human lymphatic endothelial cells (LECs) were cultured according to the manufacturer’s recommendations (Lonza, CA) and LEC proliferation was assayed with Brdu proliferation assays. For impedance assays, cells were grown in EGM-2 media with supplement. LECs were harvested, centrifuged at 1000 rpm for 5 min, counted, and resuspended in plating media. They were seeded in 96-well CellKey plates at a density of 65,000 cells/well. Plates were cultured overnight at 37°C. LECs in the 96-well CellKey plates were treated with FBS, VEGF-A, -C, -C156S and -D in the presence or absence of MAP kinase inhibitors (ERK, JNK and p38).
After VEGF-C introduction, higher impedance was observed in human LECs in a concentration-dependant manner as compared to stimulation with VEGF-A, -C156S, VEGF-D. JNK kinase inhibitors inhibited both VEGF-A and VEGF-C induced impedance, P38 kinase inhibitor had preferential inhibition of VEGF-C induced impedance, and ERK kinase inhibitors had little effect on the VEGF-C induced impedance.
The concentration-dependent changes in proliferation and impedance observed suggest VEGF-C may alter LEC proliferation, activation and barrier function. Understanding of the mechanism of VEGF-C-regulated lymphatic function may provide therapeutic intervention for the treatment of lymphangiogenesis-related disorders, including corneal transplant rejection, lymphedema, and malignancy.
This PDF is available to Subscribers Only