Skip to main content
SpringerLink
Log in

Preparation and Characterization of Circulating Angiogenic Cells for Tissue Engineering Applications

  • Protocol
  • First Online:
Cardiac Tissue Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1181))

Abstract

Circulating angiogenic cells (CACs) are a heterogeneous cell population of bone marrow (BM) origin. These cells are most commonly derived from the peripheral blood, bone marrow, and cord blood, and are one of the leading candidates for promoting vascularization in tissue engineering therapies. CACs can be isolated by culturing peripheral blood mononuclear cells (PBMCs) on fibronectin or by flow cytometry to obtain more specific subpopulations. Here we will describe how to generate a population of CACs, and how to characterize the cells and confirm their phenotype. Also, we will provide select methods that can be used to assess the angiogenic and endothelial cell-like properties of the CACs.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Asahara T, Murohara T, Sullivan A et al (1997) Isolation of putative progenitor endothelial cells for angiogenesis. Science 275:964–967

    Article  CAS  Google Scholar 

  2. Asahara T, Takahashi T, Masuda H et al (1999) VEGF contributes to postnatal neovascularization by mobilizing bone marrow-derived endothelial progenitor cells. EMBO J 18:3964–3972

    Article  CAS  Google Scholar 

  3. Grenier G, Scime A, Le Grand F et al (2007) Resident endothelial precursors in muscle, adipose, and dermis contribute to postnatal vasculogenesis. Stem Cells 25:3101–3110

    Article  CAS  Google Scholar 

  4. Tamaki T, Akatsuka A, Ando K et al (2002) Identification of myogenic-endothelial progenitor cells in the interstitial spaces of skeletal muscle. J Cell Biol 157:571–577

    Article  CAS  Google Scholar 

  5. Asahara T, Kawamoto A, Masuda H (2011) Concise review: circulating endothelial progenitor cells for vascular medicine. Stem Cells 29:1650–1655

    Article  CAS  Google Scholar 

  6. Moreno PR, Sanz J, Fuster V (2009) Promoting mechanisms of vascular health: circulating progenitor cells, angiogenesis, and reverse cholesterol transport. J Am Coll Cardiol 53:2315–2323

    Article  CAS  Google Scholar 

  7. Fadini GP, Losordo D, Dimmeler S (2012) Critical reevaluation of endothelial progenitor cell phenotypes for therapeutic and diagnostic use. Circ Res 110:624–637

    Article  CAS  Google Scholar 

  8. Gilbert PM, Blau HM (2011) Engineering a stem cell house into a home. Stem Cell Res Ther 2:3

    Article  CAS  Google Scholar 

  9. Kuraitis D, Giordano C, Ruel M et al (2012) Exploiting extracellular matrix-stem cell interactions: a review of natural materials for therapeutic muscle regeneration. Biomaterials 33:428–443

    Article  CAS  Google Scholar 

  10. Segers VF, Lee RT (2011) Biomaterials to enhance stem cell function in the heart. Circ Res 109:910–922

    Article  CAS  Google Scholar 

  11. Nunes SS, Song H, Chiang CK et al (2011) Stem cell-based cardiac tissue engineering. J Cardiovasc Transl Res 4:592–602

    Article  Google Scholar 

  12. Delewi R, Andriessen A, Tijssen JG et al (2013) Impact of intracoronary cell therapy on left ventricular function in the setting of acute myocardial infarction: a meta-analysis of randomised controlled clinical trials. Heart 99:225–232

    Article  Google Scholar 

  13. Jeevanantham V, Butler M, Saad A et al (2012) Adult bone marrow cell therapy improves survival and induces long-term improvement in cardiac parameters: a systematic review and meta-analysis. Circulation 126:551–568

    Article  Google Scholar 

  14. Amini AR, Laurencin CT, Nukavarapu SP (2012) Differential analysis of peripheral blood- and bone marrow-derived endothelial progenitor cells for enhanced vascularization in bone tissue engineering. J Orthop Res 30:1507–1515

    Article  CAS  Google Scholar 

  15. Critser PJ, Voytik-Harbin SL, Yoder MC (2011) Isolating and defining cells to engineer human blood vessels. Cell Prolif 44(Suppl 1):15–21

    Article  Google Scholar 

  16. Zhang Z, Ito WD, Hopfner U et al (2011) The role of single cell derived vascular resident endothelial progenitor cells in the enhancement of vascularization in scaffold-based skin regeneration. Biomaterials 32:4109–4117

    Article  CAS  Google Scholar 

  17. Hirschi KK, Ingram DA, Yoder MC (2008) Assessing identity, phenotype, and fate of endothelial progenitor cells. Arterioscler Thromb Vasc Biol 28:1584–1595

    Article  CAS  Google Scholar 

  18. Favre J, Terborg N, Horrevoets AJ (2013) The diverse identity of angiogenic monocytes. Eur J Clin Invest 43:100–107

    Article  Google Scholar 

  19. Kalka C, Masuda H, Takahashi T et al (2000) Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization. Proc Natl Acad Sci U S A 97:3422–3427

    Article  CAS  Google Scholar 

  20. Kuraitis D, Hou C, Zhang Y et al (2011) Ex vivo generation of a highly potent population of circulating angiogenic cells using a collagen matrix. J Mol Cell Cardiol 51:187–197

    Article  CAS  Google Scholar 

  21. Mackie AR, Losordo DW (2011) CD34-positive stem cells: in the treatment of heart and vascular disease in human beings. Tex Heart Inst J 38:474–485

    Google Scholar 

  22. Yang J, Ii M, Kamei N et al (2011) CD34+ cells represent highly functional endothelial progenitor cells in murine bone marrow. PLoS One 6:e20219

    Article  CAS  Google Scholar 

  23. Garmy-Susini B, Varner JA (2005) Circulating endothelial progenitor cells. Br J Cancer 93:855–858

    Article  CAS  Google Scholar 

  24. Peichev M, Naiyer AJ, Pereira D et al (2000) Expression of VEGFR-2 and AC133 by circulating human CD34(+) cells identifies a population of functional endothelial precursors. Blood 95:952–958

    CAS  Google Scholar 

  25. Sofrenovic T, McEwan K, Crowe S et al (2012) Circulating angiogenic cells can be derived from cryopreserved peripheral blood mononuclear cells. PLoS One 7:e48067

    Article  CAS  Google Scholar 

  26. Werner N, Kosiol S, Schiegl T et al (2005) Circulating endothelial progenitor cells and cardiovascular outcomes. N Engl J Med 353:999–1007

    Article  CAS  Google Scholar 

  27. Khan SS, Solomon MA, McCoy JP Jr (2005) Detection of circulating endothelial cells and endothelial progenitor cells by flow cytometry. Cytometry B Clin Cytom 64:1–8

    Article  Google Scholar 

  28. Shepherd RM, Capoccia BJ, Devine SM et al (2006) Angiogenic cells can be rapidly mobilized and efficiently harvested from the blood following treatment with AMD3100. Blood 108:3662–3667

    Article  CAS  Google Scholar 

  29. Steinmetz M, Nickenig G, Werner N (2010) Endothelial-regenerating cells: an expanding universe. Hypertension 55:593–599

    Article  CAS  Google Scholar 

  30. Yip HK, Chang LT, Chang WN et al (2008) Level and value of circulating endothelial progenitor cells in patients after acute ischemic stroke. Stroke 39:69–74

    Article  Google Scholar 

  31. Rafii S, Lyden D (2003) Therapeutic stem and progenitor cell transplantation for organ vascularization and regeneration. Nat Med 9:702–712

    Article  CAS  Google Scholar 

  32. Werner N, Wassmann S, Ahlers P et al (2007) Endothelial progenitor cells correlate with endothelial function in patients with coronary artery disease. Basic Res Cardiol 102:565–571

    Article  Google Scholar 

  33. Delorme B, Basire A, Gentile C et al (2005) Presence of endothelial progenitor cells, distinct from mature endothelial cells, within human CD146+ blood cells. Thromb Haemost 94:1270–1279

    CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Canadian Institutes of Health Research (to MR and EJS). BM was supported by a University of Ottawa Cardiology Research Endowment Fellowship.

Author information

Authors and Affiliations

  1. Division of Cardiac Surgery, University of Ottawa Heart Institute, 40 Ruskin Street, Ottawa, ON, Canada, K1Y 4W7

    Aleksandra Ostojic, Suzanne Crowe, Brian McNeill, Marc Ruel & Erik J. Suuronen

Authors
  1. Aleksandra Ostojic
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Suzanne Crowe
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Brian McNeill
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Marc Ruel
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Erik J. Suuronen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Erik J. Suuronen .

Editor information

Editors and Affiliations

  1. Department Chemical Engineering&Applied Chemistry, University of Toronto, Toronto, Ontario, Canada

    Milica Radisic

  2. Department of Biomedical Engineering, Tufts University, Medford, Massachusetts, USA

    Lauren D. Black III

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Ostojic, A., Crowe, S., McNeill, B., Ruel, M., Suuronen, E.J. (2014). Preparation and Characterization of Circulating Angiogenic Cells for Tissue Engineering Applications. In: Radisic, M., Black III, L. (eds) Cardiac Tissue Engineering. Methods in Molecular Biology, vol 1181. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1047-2_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-1047-2_3

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1046-5

  • Online ISBN: 978-1-4939-1047-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation