Early lymph vessel development from embryonic stem cells.
OBJECTIVE
The purpose of this study was to establish a model system for lymph vessel development based on directed differentiation of murine embryonic stem cells.


RESULTS
Stem cells were aggregated to form embryoid bodies, and subsequently cultured in 3-dimensional collagen matrix for up to 18 days. Treatment with vascular endothelial growth factor (VEGF)-C and VEGF-A individually enhanced formation of lymphatic vessel structures, although combined treatment with VEGF-C and VEGF-A was most potent and gave rise to a network of LYVE-1, podoplanin, Prox1, and VEGF receptor-3 positive lymphatic vessel structures running parallel to and apparently emanating from, capillaries. In contrast, fibroblast growth factor-2, hepatocyte growth factor, or hypoxia had little or no effect on the development of the early lymphatics. Further, cells of hematopoietic origin were shown to express lymphatic markers. In summary, different subpopulations of lymphatic endothelial cells were identified on the basis of differential expression of several lymphatic and blood vessel markers, indicating vascular heterogeneity.


CONCLUSIONS
We conclude that the present model closely mimics the early steps of lymph vessel development in mouse embryos.
Department of Genetics and Pathology, Uppsala University, Sweden. johan.kreuger@genpat.uu.se
Johan
J
Kreuger
Ingrid
I
Nilsson
Dontscho
D
Kerjaschki
Tatiana
T
Petrova
Kari
K
Alitalo
Lena
L
Claesson-Welsh
D000818:Animals; D002454:Cell Differentiation; D015687:Cell Hypoxia; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006023:Glycoproteins / Q000032:analysis / Q000502:physiology; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D018398:Homeodomain Proteins / Q000032:analysis; D042583:Lymphangiogenesis; D026901:Membrane Transport Proteins; D051379:Mice; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis; D013234:Stem Cells / Q000166:cytology*; D025521:Tumor Suppressor Proteins; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D042582:Vascular Endothelial Growth Factor C / Q000494:pharmacology; D000098551:Prospero-Related Homeobox 1 Protein
D016222:Fibroblast Growth Factor 2; D006023:Glycoproteins; D017228:Hepatocyte Growth Factor; D018398:Homeodomain Proteins; D026901:Membrane Transport Proteins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D025521:Tumor Suppressor Proteins; D042461:Vascular Endothelial Growth Factor A; D042582:Vascular Endothelial Growth Factor C; D000098551:Prospero-Related Homeobox 1 Protein; C492273:LYVE1 protein, mouse