[Immortalization of endothelial cells differentiated from mouse embryonic stem cells].
This study is designed to immortalize endothelial cells differentiated from embryonic stem cells. The embryoid bodies (EB) formed in vitro from embryonic stem cells, were induced to differentiate into many "round cells" (the precursor of endothelial cells) by retinoic acid (RA) and transforming growth factor-beta1 (TGF-beta1). These "round cells" later formed the vascular tube-like structures. Studies by scanning electronic microscopy and light microscopy and immunocytochemistry, demonstrated that these tube-like structures were constituted by a large number of round and flat cells, which were positive for "vWF" and "CD34"staining. These results indicate they are vascular endothelial cells. To immortalize these cells, human telomerase reverse transcriptase (hTERT) cDNA was transfected into "round cells" by lipofectine. hTERT mRNA expression in transfected cells was confirmed by Dot blot, RT-PCR. Furthermore, 95% these transfected cells maintain the characteristic of endothelial cell, can proliferate in large quantity in vitro, and are able to form tubular structures. These results suggest that hTERT cDNA transfection can immortalize the induced endothelial cells and therefore may provide a new source of seed cells for vascular engineering.
Shen
Gan
G
Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Shanghai 2nd Medical University, Shanghai Key Laboratory of Tissue Engineering , Shanghai 200011.
Cong
Xiao Qing
XQ
Du
Zhong Wei
ZW
Wu
Chun Fang
CF
Liu
Xiao Yin
XY
Liu
Wei
W
Cao
Yi Lin
YL
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D008853:Microscopy; D008855:Microscopy, Electron, Scanning; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology
D053773:Transforming Growth Factor beta1; D014212:Tretinoin; D019098:Telomerase