Skip to main content
SpringerLink
Log in

Expansion of Canine Bone Marrow-Derived Endothelial Progenitor Cells and Dynamic Observation

  • Basic Science Research
  • Published:
Annals of Vascular Surgery

Abstract

We cultured bone marrow-derived endothelial progenitor cells by a simple ex vivo expansion method and observed the expansion efficacy. Bone marrow mononuclear cells from five mongrel adult dogs were cultured with EGM-2MV medium in culture flasks coated with fibronectin. Morphology was observed with phase contrast microscopy, and a growth curve was constructed to evaluate the efficacy of expansion. Incorporation of Dil-ac-LDL was tested to evaluate the function. At different time points, immunocytochemical staining for flk-1, CD133, and factor VIII-related antigens was done and compared to staining of endothelial cells and mesenchymal stem cells and percentages of CD133, vascular endothelial growth factor receptor 2 (VEGFR-2), and the double-positive cells measured with flow cytometry to identify the quality and efficacy of expansion. Cluster-like attached cells grew to confluence at an average time of 10 days, and the mean number of cells harvested from 1 mL of bone marrow was (1.3 ± 0.3) × 106. The cells presented a cobblestone-like appearance and took up Dil-ac-LDL. Immunocytochemistry showed that flk-1, CD133, and factor VIII-related antigens were positive. Flow cytometry showed that VEGFR-2 and CD133 double-positive cells augmented 242-fold at the tenth day. Ex vivo expansion can effectively proliferate endothelial progenitor cells from bone marrow; the expansion efficacy could meet the requirements for tissue engineering of blood vessels.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.
Fig. 5.

Similar content being viewed by others

References

  1. Wang ZG, Du W, Li GD, et al. Rapid cellular luminal coverage of Dacron inferior vena cava prostheses in dogs by immediate seeding of antogenous endothelial cells derived from omental tissue: results of a preliminary trial. J Vasc Surg 1990;12:168–179

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  4. Gehling U, Ergun S, Schumacher U, et al. In vitro differentiation of endothelial cells from AC133 positive progenitor cells. Blood 2000;95:3106–3112

    PubMed  CAS  Google Scholar 

  5. Quirici N, Soligo D, Caneva L, et al. Differentiation and expansion of endothelial cells from human bone marrow CD133+ cells. Br J Haematol 2001;115:186–194

    Article  PubMed  CAS  Google Scholar 

  6. Boyer M, Townsend LE, Vogel M, et al. Isolation of endothelial cells and their progenitor cells from human peripheral blood. J Vasc Surg 2000;31:181–189

    Article  PubMed  CAS  Google Scholar 

  7. Seifalian AM, Salacinski HJ, Tai NR, et al. A new technique for measuring the cell growth and metabolism of endothelial cells seeded on vascular protheses. J Biomed Mater Res 2001;55:637–644

    Article  PubMed  CAS  Google Scholar 

  8. Kadner A, Hoerstrup SP, Zund G, et al. A new source for cardiovascular tissue engineering: human bone marrow stromal cells. Eur J Cardiothorac Surg 2002;21:1055–1060

    Article  PubMed  Google Scholar 

  9. Shirota T, He HB, Yasui H, et al. Human endothelial progenitor cell-seeded hybrid graft: proliferative and antithrombogenic potentials in vitro and fabrication processing. Tissue Eng 2003;9:127–135

    Article  PubMed  CAS  Google Scholar 

  10. Shintani S, Murohara T, Ikada H, et al. Augmentation of postnatal neovascularization with autologous bone marrow transplantation. Circulation 2001;103:897–903

    Article  PubMed  CAS  Google Scholar 

  11. Shi Q, Rafii S, Wu MH, et al. Evidence for circulating bone marrow-derived endothelial cells. Blood 1998;92:362–367

    PubMed  CAS  Google Scholar 

  12. Mihail H, Wolfgang E, Peter CW. Endothelial progenitor cells: mobilization, differentiation, and homing. Arterioscler Thromb Vasc Biol 2003;23:1185–1189

    Article  PubMed  CAS  Google Scholar 

  13. Moore MAS. Putting the neo into neoangiogenesis. J Clin Invest 2002;109:313–315

    Article  PubMed  CAS  Google Scholar 

  14. Moehler TM, Neben K, Ho AD, et al. Angiogenesis in hematological malignancies. Ann Hematol 2001;80:695–705

    Article  PubMed  CAS  Google Scholar 

  15. Kawamoto A, Asahara T, Losordo DW. Transplantation of endothelial progenitor cells for therapeutic neovascularization. Cardiovasc Radiat Med 2002;3:221–225

    Article  PubMed  Google Scholar 

  16. Kalka C, Masuda H, Takahashi T, et al. Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization. Proc Natl Acad Sci 2000;97:3422–3427

    Article  PubMed  CAS  Google Scholar 

  17. Stefano RD, Santoni T, Barsotti MC, et al. Different growth conditions for peripheral blood endothelial progenitors. Cardiovasc Radiat Med 2002;3:172–175

    Article  PubMed  Google Scholar 

  18. Yin AH, Miraglia S, Zanjani ED, et al. AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 1997;90:5002–5012

    PubMed  CAS  Google Scholar 

  19. Reyes M, Dudek A, Jahagirdar B, et al. Origin of endothelial progenitors in human postnatal bone marrow. J Clin Invest 2002;109:337–346

    Article  PubMed  CAS  Google Scholar 

  20. Harraz M, Jiao C, Hanlon HD, et al. CD34- blood derived human endothelial cell progenitors. Stem Cells 2001;19:304–312

    Article  PubMed  CAS  Google Scholar 

  21. Griese D, Ehsan A, Melo LG, et al. Isolation and transplantation of autologous circulating endothelial cells into denuded vessels and prothetic grafts. Circulation 2003;108:2710–2715

    Article  PubMed  Google Scholar 

  22. Wang ZG, Zhang H, Pu LQ, et al. Can endothelial seeding enhance patency and inhibit neointimal hyperplasia? Experimental studies and clinical trial of endothelial seeded venous protheses. Int Angiol 2000;19:259-269

    PubMed  CAS  Google Scholar 

Download references

Acknowledgement

We thank Dr. YeQing Cui for immunocytochemical staining and Dr. XueJing Sun for flow cytometry. We also thank Dr. YanChuan Wu for technical assistance. This work was supported by the Program of Science and Technology Committee of Beijing City under award H020920040330.

Author information

Authors and Affiliations

  1. Department of Vascular Surgery, XuanWu Hospital and the Institute of Vascular Surgery, Capital University of Medical Sciences, Beijing, People’s Republic of China

    YingFeng Wu MD, Jian Zhang MD, YongQuan Gu MD, JianXin Li PhD, Bing Chen MD, LianRui Guo MD, Tao Luo MD & ZhongGao Wang MD

  2. Department of Vascular Surgery, XuanWu Hospital and Institute of Vascular Surgery, Capital University of Medical Sciences, 45 ChangChun Street, XuanWu District, Beijing, 100053, People’s Republic of China

    YingFeng Wu MD

Authors
  1. YingFeng Wu MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jian Zhang MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. YongQuan Gu MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. JianXin Li PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bing Chen MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. LianRui Guo MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tao Luo MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. ZhongGao Wang MD
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to YingFeng Wu MD.

About this article

Cite this article

Wu, Y., Zhang, J., Gu, Y. et al. Expansion of Canine Bone Marrow-Derived Endothelial Progenitor Cells and Dynamic Observation. Ann Vasc Surg 20, 387–394 (2006). https://doi.org/10.1007/s10016-006-9047-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10016-006-9047-6

Keywords

Search

Navigation