Searching for new molecular markers for cells obtained from abdominal aortic aneurysm

Abstract

The aim of the study was to investigate specific potential markers for cells obtained from three layers of human AAA divided into three segments along the AAA based on morphological differences. The isolated cells were compared to control commercial cell types from healthy human abdominal aortas. For each type of aortic layer, three specimens from 6 patients were compared. Total RNA was isolated from 36 cell cultures for gene expression profiling and potential new cytometry markers were typed. Isolated cells were analyzed by flow cytometry by using fluorochrome-conjugated antibodies to markers: CNN1, MYH10, ENG, ICAM2, and TEK. The relative expression of 45 genes in primary cell cultures and control lines was analyzed. Statistically significant differences were found in the expression of most of the analyzed genes between individual layers and control lines. Based on relative expression, antibodies were selected for flow cytometry. Gene expression profiles allowed to select new potential cytometry markers: CNN1, MYH10, MYOCD, ENG, ICAM2, TEK. However, none of the tested markers seems to be optimal and characteristic for a specific layer of AAA.

Keywords: Abdominal aortic aneurysm (AAA); Cytometry useful markers; Gene expression; Layers of AAA.

Fig. 1

Schematic of isolation of cells specific for wall layers of AAA segments. Segments of abdominal aortic aneurysms (AAA) after fragmentation and enzyme treatment were split into 3 layers of the wall: tunica intima (internal layer—IL), tunica media (middle layer—ML) and tunica adventitia (external layer—EL)

Fig. 2

Relative expression of potential new markers. Y axis represents relative expression Nrel = 2^(ΔC normalized target − ΔC control), axis X represents expression of genes in particular cell lines

Fig. 3

Phenotypic analysis of cells by flow cytometry. Number of positive cells for the tested marker expressed in %