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Mechanisms of vasculogenesis in 3D fibrin matrices mediated by the interaction of adipose-derived stem cells and endothelial cells

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Abstract

Vascularization of tissue-engineered constructs is essential to provide sufficient nutrient supply and hemostasis after implantation into target sites. Co-cultures of adipose-derived stem cells (ASC) with outgrowth endothelial cells (OEC) in fibrin gels were shown to provide an effective possibility to induce vasculogenesis in vitro. However, the mechanisms of the interaction between these two cell types remain unclear so far. The aim of this study was to evaluate differences of direct and indirect stimulation of ASC-induced vasculogenesis, the influence of ASC on network stabilization and molecular mechanisms involved in vascular structure formation. Endothelial cells (EC) were embedded in fibrin gels either containing non-coated or ASC-coated microcarrier beads as well as ASC alone. Moreover, EC-seeded constructs incubated with ASC-conditioned medium were used in addition to constructs with ASC seeded on top. Vascular network formation was visualized by green fluorescent protein expressing cells or immunostaining for CD31 and quantified. RT-qPCR of cells derived from co-cultures in fibrin was performed to evaluate changes in the expression of EC marker genes during the first week of culture. Moreover, angiogenesis-related protein levels were measured by performing angiogenesis proteome profiler arrays. The results demonstrate that proximity of endothelial cells and ASC is required for network formation and ASC stabilize EC networks by developing pericyte characteristics. We further showed that ASC induce controlled vessel growth by secreting pro-angiogenic and regulatory proteins. This study reveals angiogenic protein profiles involved in EC/ASC interactions in fibrin matrices and confirms the usability of OEC/ASC co-cultures for autologous vascular tissue engineering.

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Acknowledgments

We thank Petra Aigner for HUVEC isolation, Andreas Teuschl for Collagen I/III and Michaela Kerschbaum for qPCR primer validation. Furthermore, we thank Philip Heher, Christiane Fuchs and Johanna Prüller (University of Applied Sciences Technikum Vienna) for their excellent technical assistance during mRNA isolation and RT-qPCR and Peter Petzelbauer (Medical University of Vienna) for his support. This project was funded by the EU Biodesign program (262948).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Donaueschingenstrasse 13, 1200, Vienna, Austria

    Sabrina Rohringer, Pablo Hofbauer, Karl H. Schneider, Anna-Maria Husa, Georg Feichtinger, Heinz Redl & Wolfgang Holnthoner

  2. Austrian Cluster for Tissue Regeneration, Vienna, Austria

    Sabrina Rohringer, Pablo Hofbauer, Karl H. Schneider, Anna-Maria Husa, Georg Feichtinger, Anja Peterbauer-Scherb, Heinz Redl & Wolfgang Holnthoner

  3. Red Cross Blood Transfusion Service, Krankenhausstrasse 7, 4020, Linz, Austria

    Anja Peterbauer-Scherb

  4. University of Applied Sciences Technikum Wien, Höchstädtplatz 6, 1200, Vienna, Austria

    Wolfgang Holnthoner

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  1. Sabrina Rohringer
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Correspondence to Wolfgang Holnthoner.

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Rohringer, S., Hofbauer, P., Schneider, K.H. et al. Mechanisms of vasculogenesis in 3D fibrin matrices mediated by the interaction of adipose-derived stem cells and endothelial cells. Angiogenesis 17, 921–933 (2014). https://doi.org/10.1007/s10456-014-9439-0

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  • DOI: https://doi.org/10.1007/s10456-014-9439-0

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