Isolation and characterization of circulating lymphatic endothelial colony forming cells

• Published in: Experimental cell research 2016-01, Vol.340 (1), p.159-169
• Main Authors: DiMaio, Terri A., Wentz, Breanna L., Lagunoff, Michael
• Format: Article
• Language: English
• Subjects: Cell Separation; Cellular biology; Endothelial Cells - cytology; Endothelial Cells - metabolism; Endothelial progenitor; Endothelium; Gene Expression Profiling; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; Humans; Lymphatic endothelial progenitor; Lymphatic system; Prox-1; Real-Time Polymerase Chain Reaction; Studies; Tumor Suppressor Proteins - genetics; Tumor Suppressor Proteins - metabolism; Vascular Endothelial Growth Factor C - genetics; Vascular Endothelial Growth Factor C - metabolism; Vascular Endothelial Growth Factor Receptor-1 - genetics; Vascular Endothelial Growth Factor Receptor-1 - metabolism; Vascular Endothelial Growth Factor Receptor-3 - genetics; Vascular Endothelial Growth Factor Receptor-3 - metabolism; VEGFR-3
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/j.yexcr.2015.11.015

The identification of circulating endothelial progenitor cells has led to speculation regarding their origin as well as their contribution to neovascular development. Two distinct types of endothelium make up the blood and lymphatic vessel system. However, it has yet to be determined whether there are distinct lymphatic-specific circulating endothelial progenitor cells. This study aims to isolate and characterize the cellular properties and global gene expression of lymphatic-specific endothelial progenitor cells. We isolated circulating endothelial colony forming cells (ECFCs) from whole peripheral blood. These cells are endothelial in nature, as defined by their expression of endothelial markers and their ability to undergo capillary morphogenesis in three-dimensional culture. A subset of isolated colonies express markers of lymphatic endothelium, including VEGFR-3 and Prox-1, with low levels of VEGFR-1, a blood endothelial marker, while the bulk of the isolated cells express high VEGFR-1 levels with low VEGFR-3 and Prox-1 expression. The different isolates have differential responses to VEGF-C, a lymphatic endothelial specific cytokine, strongly suggesting that there are lymphatic specific and blood specific ECFCs. Global analysis of gene expression revealed key differences in the regulation of pathways involved in cellular differentiation between blood and lymphatic-specific ECFCs. These data indicate that there are two distinguishable circulating ECFC types, blood and lymphatic, which are likely to have discrete functions during neovascularization. •Circulating lymphatic endothelial colony forming units exist in human blood.•Blood and lymphatic ECFCs have differential surface marker and gene expression.•Blood and lymphatic ECFCs respond differentially to VEGF-C.•ECFCs have distinct gene expression patterns from mature blood and lymphatic ECs.•Lymphatic ECFCs may play a role in lymphatic biology and disease.