Involvement of integrin-linked kinase in capillary/tube-like network formation of human vascular endothelial cells

Motomu Watanabe,Yuji Basaki,Hisayuki Kobayashi,Yayoi Fujioka-Kaneko,Michihiko Kuwano,Mamoru Kiniwa

doi:10.1251/bpo104

Abstract

Angiogenesis is a complex process involving an ECM and vascular endothelial cells (EC), and is regulated by various angiogenic factors including VEGF. The ability to form a capillary/tube-like network is a specialized function of EC. Therefore, in vitro angiogenesis was assessed by a capillary/tube-like network formation assay. There are three angiogenic parameters: capillary length, number of capillaries, and relative capillary area per field. We evaluated capillary length per field in the assay. VEGF promoted capillary/tube-like network formation of EC in a type I collagen gel matrix in vitro. Moreover, we demonstrated the involvement of ILK in a VEGF signaling pathway mediating capillary/tube-like network formation of EC using dominant-negative, kinase deficient ILK. This is a straightforward assay to monitor responses of human vascular endothelial cells.

Highlights

Angiogenesis, the formation of new blood vessels from pre-existing ones, is a critical factor in many diseases, such as diabetic retinopathy, rheumatoid arthritis, and tumor progression [1]
We investigate the effect of dominantnegative, kinase deficient integrin-linked kinase (ILK) (ILK-KD) in the capillary/tube-like network formation of human umbilical vein endothelial cells (HUVEC)
Watanabe et al - Involvement of integrin-linked kinase in capillary/tube-like network formation of human vascular endothelial cells www.biologicalprocedures.com taken using a phase-contrast microscope equipped with a CCD video camera and were evaluated by the imaging system

Summary

Introduction

Angiogenesis, the formation of new blood vessels from pre-existing ones, is a critical factor in many diseases, such as diabetic retinopathy, rheumatoid arthritis, and tumor progression [1]. VEGF promoted capillary/tube-like network formation of EC in a type I collagen gel matrix in vitro. We demonstrated the involvement of ILK in a VEGF signaling pathway mediating capillary/tube-like network formation of EC using dominant-negative, kinase deficient ILK. The ability to form a capillary/tube-like network is a specialized function of vascular endothelial cells.

Results

Conclusion