Induction of lymphatic endothelial cell differentiation in embryoid bodies

Abstract

The molecular mechanisms that regulate the formation of the lymphatic vascular system remain poorly characterized. Whereas studies in embryonic stem (ES) cells have provided major new insights into the mechanisms of blood vessel formation, the development of lymphatic endothelium has not been previously observed. We established embryoid bodies (EBs) from murine ES cells in the presence or absence of lymphangiogenic growth factors. We found that lymphatic endothelial cells develop at day 18 after EB formation. These cells express CD31 and the lymphatic lineage markers Prox-1 and Lyve-1, but not the vascular marker MECA-32, and they frequently sprout from preexisting blood vessels. Lymphatic vessel formation was potently promoted by VEGF-A and VEGF-C but not by bFGF.

Modeling lymphangiogenesis in a three-dimensional culture system

“…In addition, tube forming assays on a 3D-matrix 5 or in embryoid bodies 15 are very potent models to investigate cell migration and tube-like structure formation.…”

“…Lymphatic endothelial cell cultures are also derived from lesions induced by injecting Freund's Adjuvant into mice 5,6 . In addition, lymphatic endothelial cell differentiation are induced in embryoid bodies 15 . These systems suffer from several common drawbacks that include (1) the limited number of cells that can be obtained by isolating non-transformed cells, (2) the non-physiological features of immortalized cells, and (3) the putative dedifferentiation of cells in 2D-cultures.…”

Inhibition of endogenous TGF-β signaling enhances lymphangiogenesis

“…32,33 Briefly, R1 ES cells were trypsinized, resuspended in stem cell medium without leukemia inhibitory factor supplemented with 30 ng/mL VEGF-A at day 0. The cells were then cultured in drops hanging.…”

Sox7 and Sox17 are strain-specific modifiers of the lymphangiogenic defects caused by Sox18 dysfunction in mice

“…First, we tested the effects of lentiviral-mediated overexpression of SoxF genes in differentiating mouse ES cells, which can differentiate into endothelial cells in vitro (Balconi et al, 2000;Liersch et al, 2006). As a control, ES cells were infected with lentivirus encoding the inert marker green fluorescent protein (GFP).…”