Introduction
In vitro assays of angiogenesis face immense problems considering their reproducibility
based on the inhomogeneous characters of endothelial cells (ECs). It is necessary to detect
influencing factors, which affect the angiogenic potency of ECs.
Objective
This study aimed to analyse expression profiles of vimentin (VIM), triosephosphate isomerase
(TPI) and adenosylmethionine synthetase isoform type–2 (MAT2A) during the whole
angiogenic cascade in vitro. Furthermore, the impact of knocking down vimentin (VIM) on
angiogenesis in vitro was evaluated, while monitoring TPI and MAT2A expression.
Methods
A long–term cultivation and angiogenic stimulation of human dermal microvascular ECs
was performed. Cells were characterized via VEGFR–1 and VEGFR–2 expression and a
shRNA–mediated knockdown of VIM was performed. The process of angiogenesis in vitro
was quantified via morphological staging and mRNA–and protein–levels of all proteins were
analysed.
Results
While native cells ran through the angiogenic cascade chronologically, knockdown cells
only entered beginning stages of angiogenesis and died eventually. Cell cultures showing a
higher VEGFR–1 expression survived exclusively and displayed an upregulation of MAT2A
and TPI expression. Native cells highly expressed VIM in early stages, MAT2A mainly in the
beginning and TPI during the course of angiogenesis in vitro
