Advertisement

Angiogenesis

, Volume 21, Issue 1, pp 15–22 | Cite as

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

  • Lucas E. B. Souza
  • Liziane R. Beckenkamp
  • Lays M. Sobral
  • Daianne M. C. Fantacini
  • Fernanda U. F. Melo
  • Josiane S. Borges
  • Andréia M. Leopoldino
  • Simone Kashima
  • Dimas Tadeu Covas
Brief Communication

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.

Keywords

Angiogenesis Cell therapy Perivascular cells Adipose-derived stem cells 

Notes

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).

Compliance with ethical standards

Conflict of interest

The authors have no conflicts of interest to declare.

Supplementary material

10456_2017_9579_MOESM1_ESM.docx (5.3 mb)
Supplementary material 1 (DOCX 5376 kb)

References

  1. 1.
    Sacchetti B, Funari A, Michienzi S et al (2007) Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment. Cell 131:324–336CrossRefPubMedGoogle Scholar
  2. 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 25:1843–1852CrossRefPubMedGoogle Scholar
  3. 3.
    Zuk PA, Zhu M, Ashjian P et al (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13:4279–4295CrossRefPubMedPubMedCentralGoogle Scholar
  4. 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 20:419–427CrossRefGoogle Scholar
  5. 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 15:443–455CrossRefPubMedPubMedCentralGoogle Scholar
  6. 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 7:5CrossRefPubMedPubMedCentralGoogle Scholar
  7. 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 117:229–233CrossRefPubMedPubMedCentralGoogle Scholar
  8. 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 385:117–171CrossRefGoogle Scholar
  9. 9.
    Trounson A, McDonald C (2015) Stem cell therapies in clinical trials: progress and challenges. Cell Stem Cell 17:11–22CrossRefPubMedGoogle Scholar
  10. 10.
    Karantalis V, Hare JM (2015) Use of mesenchymal stem cells for therapy of cardiac disease. Circ Res 116:1413–1430CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Eming SA, Martin P, Tomic-Canic M (2014) Wound repair and regeneration: mechanisms, signaling, and translation. Sci Transl Med 6:265rs6CrossRefGoogle Scholar
  12. 12.
    Locatelli P, Olea FD, Hnatiuk A et al (2015) Mesenchymal stromal cells overexpressing vascular endothelial growth factor in ovine myocardial infarction. Gene Ther 22:449–457CrossRefPubMedGoogle Scholar
  13. 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 135:799–808CrossRefPubMedGoogle Scholar
  14. 14.
    Han J, Lee J-E, Jin J et al (2011) The spatiotemporal development of adipose tissue. Development 138:5027–5037CrossRefPubMedGoogle Scholar
  15. 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 119:629–641CrossRefPubMedGoogle Scholar
  16. 16.
    Suga H, Shigeura T, Matsumoto D et al (2007) Rapid expansion of human adipose-derived stromal cells preserving multipotency. Cytotherapy 9:738–745CrossRefPubMedGoogle Scholar
  17. 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 85:929–940PubMedGoogle Scholar
  18. 18.
    Schellenberg A, Stiehl T, Horn P et al (2012) Population dynamics of mesenchymal stromal cells during culture expansion. Cytotherapy 14:401–411CrossRefPubMedGoogle Scholar
  19. 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 35:640–652CrossRefPubMedGoogle Scholar
  20. 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 107:3317–3322CrossRefPubMedGoogle Scholar
  21. 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 31:117–125CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  • Lucas E. B. Souza
    • 1
    • 2
  • Liziane R. Beckenkamp
    • 1
    • 2
  • Lays M. Sobral
    • 3
  • Daianne M. C. Fantacini
    • 1
    • 2
  • Fernanda U. F. Melo
    • 2
  • Josiane S. Borges
    • 2
  • Andréia M. Leopoldino
    • 3
  • Simone Kashima
    • 2
  • Dimas Tadeu Covas
    • 1
    • 2
  1. 1.Department of Clinical Medicine, Ribeirão Preto Medical SchoolUniversity of São PauloRibeirão PretoBrazil
  2. 2.National Institute of Science and Technology in Stem Cells and Cell TherapyRibeirão PretoBrazil
  3. 3.Department of Clinical Analyses, Toxicology and Food Science, School of Pharmaceutical Sciences of Ribeirão PretoUniversity of São PauloRibeirão PretoBrazil

Personalised recommendations