Phenotypic and functional analysis of organ specific endothelial cells

• Main Author: Hidden, Sophie Kate
• Published: University of Birmingham 2010
• Subjects: 616.079; RB Pathology
• Online Access: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.525775

All classes of leukocytes must be able to move from the circulation into tissue to carry out their protective functions. To achieve this transfer, the flowing cells must adhere to the endothelium and migrate through the vessel wall. Though this process follows common stages during recruitment in different organs, there is also specialisation in underlying molecular mechanisms. The main aim was to further elucidate these organ specific phenotypic differences and relate them to the functional ability of EC to recruit subsets and total leukocytes from flow. Murine models of human disease are an incredibly useful experimental tool allowing investigation of whole diseases to the effect of one gene on a disease outcome. Isolation of primary human endothelial cell populations is well-defined (hLSEC/HUVEC) however obtaining and culturing the murine counterparts is more challenging. Primary mLSEC were isolated from murine livers using ɑ-CD146 magnetic beads and the phenotype compared to immortalised cell lines from heart (mUCEC-1), skin (s.END) and brain (b.End.5). The expression of a number of adhesion molecules and endothelial markers varied within cell type in response to pro-inflammatory insult (Endoglin, CD34, CD31). Some artefacts of immortalisation were also apparent (VCAM-1 expression in mUCEC-1 and LYVE-1 in s.END). Functional assays indicated small differences in cell types and further microarray analysis elucidated further candidates, including chemokines, which could be involved in regulating leukocyte recruitment processes in the different organs examined.