title: Efficient differentiation of human pluripotent stem cells to endothelial progenitors via small-molecule activation of WNT signaling.
abstract: Human pluripotent stem cell (hPSC)-derived endothelial cells and their progenitors may provide the means for vascularization of tissue-engineered constructs and can serve as models to study vascular development and disease. Here, we report a method to efficiently produce endothelial cells from hPSCs via GSK3 inhibition and culture in defined media to direct hPSC differentiation to CD34(+)CD31(+) endothelial progenitors. Exogenous vascular endothelial growth factor (VEGF) treatment was dispensable, and endothelial progenitor differentiation was β-catenin dependent. Furthermore, by clonal analysis, we showed that CD34(+)CD31(+)CD117(+)TIE-2(+) endothelial progenitors were multipotent, capable of differentiating into calponin-expressing smooth muscle cells and CD31(+)CD144(+)vWF(+)I-CAM1(+) endothelial cells. These endothelial cells were capable of 20 population doublings, formed tube-like structures, imported acetylated low-density lipoprotein, and maintained a dynamic barrier function. This study provides a rapid and efficient method for production of hPSC-derived endothelial progenitors and endothelial cells and identifies WNT/β-catenin signaling as a primary regulator for generating vascular cells from hPSCs.
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Lian
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Bao
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Abraham
authors.initials: A
authors.lastname: Al-Ahmad
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Jialu
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Yue
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Wentao
authors.initials: W
authors.lastname: Dong
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Kaitlin K
authors.initials: KK
authors.lastname: Dunn
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Eric V
authors.initials: EV
authors.lastname: Shusta
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA. Electronic address: palecek@engr.wisc.edu.
authors.forename: Sean P
authors.initials: SP
authors.lastname: Palecek
mesh_terms: D000631:Aminopyridines / Q000494:pharmacology; D018952:Antigens, CD34 / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002454:Cell Differentiation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D066026:Endothelial Progenitor Cells / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D015870:Gene Expression; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D011743:Pyrimidines / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism
chemical_list: D000631:Aminopyridines; D018952:Antigens, CD34; C473710:Chir 98014; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011743:Pyrimidines; D042461:Vascular Endothelial Growth Factor A; D051176:beta Catenin; D038362:Glycogen Synthase Kinase 3