Skip to main content

Advertisement

SpringerLink
Log in

Pre-culture in endothelial growth medium enhances the angiogenic properties of adipose-derived stem/stromal cells

  • Brief Communication
  • Published:
Angiogenesis Aims and scope Submit manuscript

Abstract

Considerable progress has been made on the development of adipose-derived stem/stromal cells (ASCs) as pro-angiogenic therapeutic tools. However, variable clinical results highlight the need for devising strategies to enhance their therapeutic efficacy. Since ASCs proliferate and stabilize newly formed vessels during the angiogenic phase of adipose tissue formation, we hypothesized that mimicking an angiogenic milieu during culture of ASCs would enhance their capacity to support endothelial cell survival and angiogenesis. To test this, we compared the effect of an endothelial growth medium (EGM-2) and conventional media (αMEM) on the progenitor and angiogenic properties of ASCs. ASCs cultured in EGM-2 (ASC-EGM) displayed the highest clonogenic efficiency, proliferative potential and multilineage potential. After co-culture under growth factor starvation, only ASC-EGM attenuated luciferase-expressing human umbilical vein endothelial cells (HUVECluc) apoptosis and supported the formation of endothelial cords in a dose-dependent manner. These effects were recapitulated by the conditioned medium of ASC-EGM, which displayed a 100-fold higher expression of hepatocyte growth factor in comparison with ASC-αMEM. Next, HUVECluc and ASCs were co-transplanted subcutaneously into immunodeficient mice, and the survival of HUVECluc was monitored by bioluminescent imaging. After 60 days, the survival of HUVECluc transplanted alone was equivalent to that of HUVECluc co-transplanted with ASC-αMEM (15.0 ± 0.7 vs. 13.0 ± 0.5%). Strikingly, co-transplantation with ASC-EGM increased HUVECluc survival to 105.0 ± 3.5%, and the resulting organoids displayed functional vasculature with the highest human-derived vascular area. These findings demonstrate that pre-conditioning of ASCs in endothelial growth medium augment their pro-angiogenic properties and could enhance their therapeutic efficacy against ischemic diseases.

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

References

  1. Sacchetti B, Funari A, Michienzi S et al (2007) Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment. Cell 131:324–336

    Article  CAS  PubMed  Google Scholar 

  2. Souza LEB, Malta TM, Kashima Haddad S, Covas DT (2016) Mesenchymal stem cells and pericytes: to what extent are they related? Stem Cells Dev 25:1843–1852

    Article  PubMed  Google Scholar 

  3. Zuk PA, Zhu M, Ashjian P et al (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13:4279–4295

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. da Silva Meirelles L, Fontes AM, Covas DT, Caplan AI (2009) Mechanisms involved in the therapeutic properties of mesenchymal stem cells. Cytokine Growth Factor Rev 20:419–427

    Article  Google Scholar 

  5. Lin RZ, Moreno-Luna R, Zhou B et al (2012) Equal modulation of endothelial cell function by four distinct tissue-specific mesenchymal stem cells. Angiogenesis 15:443–455

    Article  PubMed  PubMed Central  Google Scholar 

  6. Freiman A, Shandalov Y, Rozenfeld D et al (2016) Adipose-derived endothelial and mesenchymal stem cells enhance vascular network formation on three-dimensional constructs in vitro. Stem Cell Res Ther 7:5

    Article  PubMed  PubMed Central  Google Scholar 

  7. Sanina C, Hare JM (2015) Mesenchymal stem cells as a biological drug for heart disease: where are we with cardiac cell-based therapy? Circ Res 117:229–233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. GBD 2015 Mortality and Causes of Death Collaborators (2015) Global, regional, and national age-sex specific all-cause and cause-specific mortality for 240 causes of death, 1990–2013: a systematic analysis for the Global Burden of Disease Study 2013. Lancet 385:117–171

    Article  Google Scholar 

  9. Trounson A, McDonald C (2015) Stem cell therapies in clinical trials: progress and challenges. Cell Stem Cell 17:11–22

    Article  CAS  PubMed  Google Scholar 

  10. Karantalis V, Hare JM (2015) Use of mesenchymal stem cells for therapy of cardiac disease. Circ Res 116:1413–1430

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Eming SA, Martin P, Tomic-Canic M (2014) Wound repair and regeneration: mechanisms, signaling, and translation. Sci Transl Med 6:265rs6

    Article  Google Scholar 

  12. Locatelli P, Olea FD, Hnatiuk A et al (2015) Mesenchymal stromal cells overexpressing vascular endothelial growth factor in ovine myocardial infarction. Gene Ther 22:449–457

    Article  CAS  PubMed  Google Scholar 

  13. Hu X, Yu SP, Fraser JL et al (2008) Transplantation of hypoxia-preconditioned mesenchymal stem cells improves infarcted heart function via enhanced survival of implanted cells and angiogenesis. J Thorac Cardiovasc Surg 135:799–808

    Article  CAS  PubMed  Google Scholar 

  14. Han J, Lee J-E, Jin J et al (2011) The spatiotemporal development of adipose tissue. Development 138:5027–5037

    Article  CAS  PubMed  Google Scholar 

  15. Bussolino F, Di Renzo MF, Ziche M et al (1992) Hepatocyte growth factor is a potent angiogenic factor which stimulates endothelial cell motility and growth. J Cell Biol 119:629–641

    Article  CAS  PubMed  Google Scholar 

  16. Suga H, Shigeura T, Matsumoto D et al (2007) Rapid expansion of human adipose-derived stromal cells preserving multipotency. Cytotherapy 9:738–745

    Article  CAS  PubMed  Google Scholar 

  17. Gronthos S, Simmons PJ (1995) The growth factor requirements of STRO-1-positive human bone marrow stromal precursors under serum-deprived conditions in vitro. Blood 85:929–940

    CAS  PubMed  Google Scholar 

  18. Schellenberg A, Stiehl T, Horn P et al (2012) Population dynamics of mesenchymal stromal cells during culture expansion. Cytotherapy 14:401–411

    Article  CAS  PubMed  Google Scholar 

  19. Zwezdaryk KJ, Coffelt SB, Figueroa YG et al (2007) Erythropoietin, a hypoxia-regulated factor, elicits a pro-angiogenic program in human mesenchymal stem cells. Exp Hematol 35:640–652

    Article  CAS  PubMed  Google Scholar 

  20. Yang F, Cho S-W, Son SM et al (2010) Genetic engineering of human stem cells for enhanced angiogenesis using biodegradable polymeric nanoparticles. Proc Natl Acad Sci USA 107:3317–3322

    Article  CAS  PubMed  Google Scholar 

  21. Zhu X-Y, Urbieta-Caceres V, Krier JD et al (2013) Mesenchymal stem cells and endothelial progenitor cells decrease renal injury in experimental swine renal artery stenosis through different mechanisms. Stem Cells 31:117–125

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Dr. Marina Rosique for providing us the samples of human adipose tissue as well as Patrícia Palma and Camila Menezes for their support with the acquisition of flow cytometry data. This work was funded with resources from São Paulo Research Foundation (FAPESP, Brazil), Coordination for Improvement of Higher Education Personnel (CAPES, Brazil) and from the National Counsel of Technological and Scientific Development (CNPq, Brazil).

Author information

Authors and Affiliations

  1. Department of Clinical Medicine, Ribeirão Preto Medical School, University of São Paulo, 3900 Bandeirantes Avenue, Ribeirão Preto, São Paulo, 14048-900, Brazil

    Lucas E. B. Souza, Liziane R. Beckenkamp, Daianne M. C. Fantacini & Dimas Tadeu Covas

  2. National Institute of Science and Technology in Stem Cells and Cell Therapy, Ribeirão Preto, São Paulo, Brazil

    Lucas E. B. Souza, Liziane R. Beckenkamp, Daianne M. C. Fantacini, Fernanda U. F. Melo, Josiane S. Borges, Simone Kashima & Dimas Tadeu Covas

  3. Department of Clinical Analyses, Toxicology and Food Science, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Café Avenue, Ribeirão Preto, São Paulo, 14040-903, Brazil

    Lays M. Sobral & Andréia M. Leopoldino

Authors
  1. Lucas E. B. Souza
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Liziane R. Beckenkamp
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lays M. Sobral
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Daianne M. C. Fantacini
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Fernanda U. F. Melo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Josiane S. Borges
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Andréia M. Leopoldino
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Simone Kashima
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Dimas Tadeu Covas
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lucas E. B. Souza.

Ethics declarations

Conflict of interest

The authors have no conflicts of interest to declare.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 5376 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Souza, L.E.B., Beckenkamp, L.R., Sobral, L.M. et al. Pre-culture in endothelial growth medium enhances the angiogenic properties of adipose-derived stem/stromal cells. Angiogenesis 21, 15–22 (2018). https://doi.org/10.1007/s10456-017-9579-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10456-017-9579-0

Keywords

Search

Navigation