title: Low oxygen tension enhances endothelial fate of human pluripotent stem cells.
abstract: OBJECTIVE
  A critical regulator of the developing or regenerating vasculature is low oxygen tension. Precise elucidation of the role of low oxygen environments on endothelial commitment from human pluripotent stem cells necessitates controlled in vitro differentiation environments.
  
  
  RESULTS
  We used a feeder-free, 2-dimensional differentiation system in which we could monitor accurately dissolved oxygen levels during human pluripotent stem cell differentiation toward early vascular cells (EVCs). We found that oxygen uptake rate of differentiating human pluripotent stem cells is lower in 5% O2 compared with atmospheric conditions. EVCs differentiated in 5% O2 had an increased vascular endothelial cadherin expression with clusters of vascular endothelial cadherin+ cells surrounded by platelet-derived growth factor β+ cells. When we assessed the temporal effects of low oxygen differentiation environments, we determined that low oxygen environments during the early stages of EVC differentiation enhance endothelial lineage commitment. EVCs differentiated in 5% O2 exhibited an increased expression of vascular endothelial cadherin and CD31 along with their localization to the membrane, enhanced lectin binding and acetylated low-density lipoprotein uptake, rapid cord-like structure formation, and increased expression of arterial endothelial cell markers. Inhibition of reactive oxygen species generation during the early stages of differentiation abrogated the endothelial inductive effects of the low oxygen environments.
  
  
  CONCLUSIONS
  Low oxygen tension during early stages of EVC derivation induces endothelial commitment and maturation through the accumulation of reactive oxygen species, highlighting the importance of regulating oxygen tensions during human pluripotent stem cell-vascular differentiation.
authors: From the Department of Chemical and Biomolecular Engineering, Johns Hopkins Physical Sciences-Oncology Center, Institute for NanoBioTechnology (S.K., P.P., S.G.), Department of Biomedical Engineering (S.K.), Cellular and Molecular Biology (E.P.), and Department of Materials Science and Engineering (S.G.), Johns Hopkins University, Baltimore, MD.
authors: Sravanti
authors: S
authors: Kusuma
authors: Elizabeth
authors: E
authors: Peijnenburg
authors: Parth
authors: P
authors: Patel
authors: Sharon
authors: S
authors: Gerecht
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D002454:Cell Differentiation; D015687:Cell Hypoxia; D002460:Cell Line; D019070:Cell Lineage; D002462:Cell Membrane / Q000378:metabolism; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000378:metabolism*; D061252:Feeder Cells; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D021381:Protein Transport; D017382:Reactive Oxygen Species / Q000378:metabolism; D055153:Stem Cell Niche; D013997:Time Factors; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015415:Biomarkers; D015820:Cadherins; D037102:Lectins; D008077:Lipoproteins, LDL; D010100:Oxygen; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D017382:Reactive Oxygen Species; D000099152:Cadherin 5; C032629:acetyl-LDL
keywords: endothelial cells; markers, differentiation; oxygen; pluripotent stem cells