Vasculogenesis from Embryonic Bodies of Murine Embryonic Stem Cells Transfected by Tgf-β1 Gene

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Abstract
Mouse embryonic stem (ES) cells transfected with a 1.7 kb cDNA of porcine transforming growth factor type beta1 (TGFbeta1), known as ES-T cells, were found to be able to differentiate in vitro into cystic embryonic bodies (EBs) with outspread tubular structures. Morphological analysis using light, phase-contrast and electron microscopes revealed that in culture, the EBs of ES-T cells initially developed some flat endothelial-like cells which further proliferated and migrated to form thread structures. At 8-10 days after EB formation, these thread structures further developed into net-like and tubular structures connecting directly to EBs. Immunofluorescent assays using antibodies against Flk-1 and von Willebrand factor (vWF) indicated that these net-like and tubular structures of ES-T cells consisted of vascular endothelial cells. Further analysis by RT-PCR revealed that the EBs with tubular structures expressed the mRNA of other markers of vascular endothelial cells, including VE-cadherin and platelet-endothelial cell adhesion molecule (PECAM). Cells of hematopoietic origin were not detected on the outside of EBs by immunostaining using several antibodies specific for granulocytes, macrophages and lymphocytes as well as by benzidine staining for erythroid cells on the outside of EBs. Our data demonstrates that the transfer of TGFbeta1 into ES cells results in a significant vasculogenesis without concomitant hematopoiesis. ES-T cells could therefore provide an excellent model for studying blood vessel formation and vasculogenic and hematopoietic interactions.

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