Spontaneous endothelial-to-mesenchymal transition in human primary umbilical vein endothelial cells

Highlights . Spontaneous endothelial-to-mesenchymal transition of primary human umbilical vein endothelial cells (HUVEC) is characterized by an acquired expression SNAI2 and TWIST1 genes, loss markers transcription factors (CD31/PECAM1, VE-cadherin, ERG factor), pronounced S100A4 ACTA2 active production type I collagen, a major component the extracellular matrix. An optimal algorithm to detect includes gene profiling lineage ( PECAM1, CDH5, VWF, ), factors, mesenchymal specification FAP, S100A4, ) matrix synthesis COL1A1, COL1A2 along with subsequent negative staining for CD31/PECAM1, or positive intracellular collagen. Aim To develop tools determine (EndoMT) in vitro Methods We examined two batches where first cell batch had conventional morphology second underwent spontaneous EndoMT. Human coronary artery (HCAEC) internal thoracic (HITAEC) were used as control Molecular profile was assessed means reverse transcription-quantitative polymerase chain reaction, Western blotting, immunofluorescence further confocal microscopy. Results In contrast HUVEC physiological arterial ECs, undergoing EndoMT lost SNAI2, components while retaining common vascular HES1, NRP1 ). blotting analysis confirmed VE-cadherin/CDH5, ERG) demonstrated retained abovementioned markers. Negligible MYH11 SMTN genes encoding specific contractile (smooth muscle myosin heavy smoothelin) combination less marker alpha smooth actin indicated phenotypic identity EndoMT-transformed myofibroblasts but not cells. Loss (CD31/PECAM-1, factor) collagen testified ongoing Conclusion assess implies measurement ERG, TWIST1, ACTA2, COL1A1 , respective CD31/PECAM-1, factor