Journal Mobile Options
Table of Contents
Vol. 181, No. 2, 2005
Issue release date: March 2006

Development and Characterization of a Spheroidal Coculture Model of Endothelial Cells and Fibroblasts for Improving Angiogenesis in Tissue Engineering

Wenger A. · Kowalewski N. · Stahl A. · Mehlhorn A.T. · Schmal H. · Stark G.B. · Finkenzeller G.
To view the fulltext, log in and/or choose pay-per-view option

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


Neovascularization is a critical step in tissue engineering applications since implantation of voluminous grafts without sufficient vascularity results in hypoxic cell death of central tissues. We have developed a three-dimensional spheroidal coculture system consisting of human umbilical vein endothelial cells (HUVECs) and human primary fibroblasts (hFBs) to improve angiogenesis in tissue engineering applications. Morphological analysis of cryosections from HUVEC/hFB cospheroids revealed a characteristic temporal and spatial organization with HUVECs located in the center of the cospheroid and a peripheral localization of fibroblasts. In coculture spheroids, the level of apoptosis of endothelial cells was strongly decreased upon cocultivation with fibroblasts. Collagen-embedded HUVEC spheroids develop numerous lumenized capillary-like sprouts. This was also apparent for HUVEC/hFB cospheroids, albeit to a lesser extent. Quantification of cumulative sprout length revealed an approximately 35% reduction in endothelial cell sprouting upon cocultivation with fibroblasts in cospheroids. The slight reduction in endothelial cell sprouting was not mediated by a paracrine mechanism but is most likely due to the formation of heterogenic cell contacts between HUVECs and hFBs within the cospheroid. The model system introduced in this study is suitable for the development of a preformed lumenized capillary-like network ex vivo and may therefore be useful for improving angiogenesis in in vivo tissue engineering applications.

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Zisch, A.H., M.P. Lutolf, J.A. Hubbell (2003) Biopolymeric delivery matrices for angiogenic growth factors. Cardiovasc Pathol 12: 295–310.
  2. Wenger, A., A. Stahl, H. Weber, G. Finkenzeller, H.G. Augustin, G.B. Stark, U. Kneser (2004) Modulation of in vitro angiogenesis in a three-dimensional spheroidal coculture model for bone tissue engineering. Tissue Eng 10: 1536–1547.
  3. Sieminski, A.L, R.F. Padera, T. Blunk, K.J. Gooch (2002) Systemic delivery of human growth hormone using genetically modified tissue-engineered microvascular networks: prolonged delivery and endothelial survival with inclusion of nonendothelial cells. Tissue Eng 8: 1057–1069.
  4. Schumacher, B., P. Pecher, B.U. von Specht, T. Stegmann (1998) Induction of neoangiogenesis in ischemic myocardium by human growth factors: first clinical results of a new treatment of coronary heart disease. Circulation 97: 645–650.
  5. Schechner, J.S., A.K. Nath, L. Zheng, M.S. Kluger, C.C. Hughes, M.R. Sierra-Honigmann, M.I. Lorber, G. Tellides, M. Kashgarian, A.L. Bothwell, J.S. Pober (2000) In vivo formation of complex microvessels lined by human endothelial cells in an immunodeficient mouse. Proc Natl Acad Sci USA 97: 9191–9196.
  6. Rebar, E.J., Y. Huang, R. Hickey, A.K. Nath, D. Meoli, S. Nath, B. Chen, L. Xu, Y. Liang, A.C. Jamieson, L. Zhang, S.K. Spratt, C.C. Case, A. Wolffe, F.J. Giordano (2002) Induction of angiogenesis in a mouse model using engineered transcription factors. Nat Med 8: 1427–1432.
  7. Pieper, J.S., T. Hafmans, P.B. van Wachem, M.J. van Luyn, L.A. Brouwer, J.H. Veerkamp, T.H. van Kuppevelt (2002) Loading of collagen-heparan sulfate matrices with bFGF promotes angiogenesis and tissue generation in rats. J Biomed Mater Res 62: 185–194.
  8. Perets, A., Y. Baruch, F. Weisbuch, G. Shoshany, G. Neufeld, S. Cohen (2003) Enhancing the vascularization of three-dimensional porous alginate scaffolds by incorporating controlled release basic fibroblast growth factor microspheres. J Biomed Mater Res A 65: 489–497.

    External Resources

  9. Neumann, T., B.S. Nicholson, J.E. Sanders (2003) Tissue engineering of perfused microvessels. Microvasc Res 66: 59–67.
  10. Korff, T., S. Kimmina, G. Martiny-Baron, H.G. Augustin (2001) Blood vessel maturation in a 3-dimensional spheroidal coculture model: direct contact with smooth muscle cells regulates endothelial cell quiescence and abrogates VEGF responsiveness. FASEB J 15: 447–457.
  11. Korff, T., H.G. Augustin (1999) Tensional forces in fibrillar extracellular matrices control directional capillary sprouting. J Cell Sci 112: 3249–3258.
  12. Korff, T., H.G. Augustin (1998) Integration of endothelial cells in multicellular spheroids prevents apoptosis and induces differentiation. J Cell Biol 143: 1341–1352.
  13. Kipshidze, N., V. Chekanov, P. Chawla, L.R. Shankar, J.B. Gosset, K. Kumar, D. Hammen, J. Gordon, M.H. Keelan (2000) Angiogenesis in a patient with ischemic limb induced by intramuscular injection of vascular endothelial growth factor and fibrin platform. Tex Heart Inst J 27: 196–200.
  14. Henry, T.D., K. Rocha-Singh, J.M. Isner, D.J. Kereiakes, F.J. Giordano, M. Simons, D.W. Losordo, R.C. Hendel, R.O. Bonow, S.M. Eppler, T.F. Zioncheck, E.B. Holmgren, E.R. McCluskey (2001) Intracoronary administration of recombinant human vascular endothelial growth factor to patients with coronary artery disease. Am Heart J 142: 872–880.
  15. Haspel, H.C., G.M. Scicli, G. McMahon, A.G. Scicli (2002) Inhibition of vascular endothelial growth factor-associated tyrosine kinase activity with SU5416 blocks sprouting in the microvascular endothelial cell spheroid model of angiogenesis. Microvasc Res 63: 304–315.
  16. Griffith C.K., C. Miller, R.C. Sainson, J.W. Calvert, N.L. Jeon, C.C. Hughes, S.C. George (2005) Diffusion limits of an in vitro thick prevascularized tissue. Tissue Eng 11: 257–266.
  17. Enis, D.R., B.R. Shepherd, Y. Wang, A. Qasim, C.M. Shanahan, P.L. Weissberg, M. Kashgarian, J.S. Pober, J.S. Schechner (2005) Induction, differentiation, and remodeling of blood vessels after transplantation of Bcl-2-transduced endothelial cells. Proc Natl Acad Sci USA 102: 425–430.
  18. Dor, Y., V. Djonov, E. Keshet (2003) Induction of vascular networks in adult organs: implications to proangiogenic therapy. Ann N Y Acad Sci 995: 208–216.
  19. Delia, D., M.G. Lampugnani, M. Resnati, E. Dejana, A. Aiello, E. Fontanella, D. Soligo, M.A. Pierotti, M.F. Greaves (1993) CD34 expression is regulated reciprocally with adhesion molecules in vascular endothelial cells in vitro. Blood 81: 1001–1008.
  20. Bouis, D., M.C. Boelens, E. Peters, P. Koolwijk, G. Stob, I.P. Kema, M. Klinkenberg, N.H. Mulder, G.A. Hospers (2003) Combination of vascular endothelial growth factor (VEGF) and thymidine phosphorylase (TP) to improve angiogenic gene therapy. Angiogenesis 6: 185–192.
  21. Benjamin, L.E., I. Hemo, E. Keshet (1998) A plasticity window for blood vessel remodelling is defined by pericyte coverage of the preformed endothelial network and is regulated by PDGF-B and VEGF. Development 125: 1591–1598.
  22. Baumgartner, I., A. Pieczek, O. Manor, R. Blair, M. Kearney, K. Walsh, J.M. Isner (1998) Constitutive expression of phVEGF165 after intramuscular gene transfer promotes collateral vessel development in patients with critical limb ischemia. Circulation 97: 1114–1123.

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50