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In vitro vessel-forming capacity of endothelial progenitor cells in high glucose conditions

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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.

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Acknowledgment

This study was supported by grants from the Thailand Research Fund (grant no. RTA 488–0007), the Commission on Higher Education (grant no. CHE-RES-RG-49), and the Siriraj Thesis Graduate Scholarship. S. Issaragrisil is a senior research scholar of Thailand Research Fund.

Disclosures

All authors have no conflicts of interest.

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Authors and Affiliations

  1. Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand

    Chaiwat Jiraritthamrong

  2. Division of Cell Biology, Department of Pre-clinical Sciences, Faculty of Medicine, Thammasat University, Khlong Luang, Pathumthani, Thailand

    Pakpoom Kheolamai, Sirikul Manochantr & Chairat Tantrawatpan

  3. Division of Hematology, Department of Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand

    Yaowalak U-Pratya & Surapol Issaragrisil

  4. Siriraj Center of Excellence for Stem Cell Research, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand

    Methichit Chayosumrit, Hathaitip Sritanaudomchai & Surapol Issaragrisil

  5. Department of Clinical Microscopy, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand

    Aungkura Supokawej

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  1. Chaiwat Jiraritthamrong
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Correspondence to Surapol Issaragrisil.

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Jiraritthamrong, C., Kheolamai, P., U-Pratya, Y. et al. In vitro vessel-forming capacity of endothelial progenitor cells in high glucose conditions. Ann Hematol 91, 311–320 (2012). https://doi.org/10.1007/s00277-011-1300-6

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  • DOI: https://doi.org/10.1007/s00277-011-1300-6

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