title: In vitro vessel-forming capacity of endothelial progenitor cells in high glucose conditions.
abstract: In type 2 diabetes, the impairment of vascular repair processes and angiogenesis are due to endothelial progenitor cell (EPC) dysfunction. In this study, we established a quantitative methodology to assess EPC function by using an in vitro 5-(6)-carboxyfluorescein diacetate succinimidyl ester-labeling vessel formation assay. The EPCs were cultured in three different glucose concentrations (100, 189.5, and 295.5 mg/dl of D: -glucose) representing normal control and diabetes with either good or poor glycemic control, respectively. We found that the in vitro vessel-forming capacity was impaired in EPCs cultured in high glucose concentrations compared to normal control (43.4 ± 0.8% and 34.7 ± 0.7% vs. 50.8 ± 2.1%). We further studied expression of various genes involved in vessel formation. There was a lower level of angiopoietin 1 gene expression in EPCs cultured in high glucose concentrations. The addition of recombinant angiopoietin 1 significantly increased the vessel-forming capacity of EPCs cultured in high glucose concentration (35.3 ± 2.0% to 48.8 ± 2.7%), whereas the addition of angiopoietin 2 (a competitive inhibitor of angiopoietin 1) impaired the vessel-forming capacity of EPCs cultured in normal glucose concentration (51.8 ± 1.3% to 41.3 ± 0.6%). We conclude that the in vitro vessel-forming capacity of EPCs cultured in high glucose concentration is impaired due to low levels of angiopoietin 1.
authors.affiliation: Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Chaiwat
authors.initials: C
authors.lastname: Jiraritthamrong
authors.forename: Pakpoom
authors.initials: P
authors.lastname: Kheolamai
authors.forename: Yaowalak
authors.initials: Y
authors.lastname: U-Pratya
authors.forename: Methichit
authors.initials: M
authors.lastname: Chayosumrit
authors.forename: Aungkura
authors.initials: A
authors.lastname: Supokawej
authors.forename: Sirikul
authors.initials: S
authors.lastname: Manochantr
authors.forename: Chairat
authors.initials: C
authors.lastname: Tantrawatpan
authors.forename: Hathaitip
authors.initials: H
authors.lastname: Sritanaudomchai
authors.forename: Surapol
authors.initials: S
authors.lastname: Issaragrisil
mesh_terms: D042683:Angiopoietin-1 / Q000494:pharmacology; D042702:Angiopoietin-2 / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003924:Diabetes Mellitus, Type 2 / Q000503:physiopathology; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology*; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D020869:Gene Expression Profiling; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D018919:Neovascularization, Physiologic / Q000187:drug effects* / Q000502:physiology; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D013388:Succinimides / Q000378:metabolism
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D042683:Angiopoietin-1; D042702:Angiopoietin-2; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013388:Succinimides; D005947:Glucose