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Porous Membranes Promote Endothelial Differentiation of Adipose-Derived Stem Cells and Perivascular Interactions

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Abstract

Efforts to create physiologically relevant microenvironments for cell differentiation have led to the creation of three-dimensional (3D) support matrices with varying physical and chemical properties. In an effort to simplify the complexity of these matrices, while maintaining their unique permeable nature, we investigated the culture and differentiation of adipose-derived stem cells (ADSCs) on porous membranes. Membranes offer many of the benefits of a 3D matrix, but simplify cell seeding, imaging, analysis and post-differentiation recovery. After inducing the differentiation of ADSCs into endothelial cells (ECs), the cells cultured on porous substrates produce more branch points and greater tube length in angiogenesis assays compared to the cells cultured on non-porous controls. While we confirm that ADSCs can be induced with vascular endothelial growth factor to express endothelial adhesion molecule CD31 (PECAM-1), only when co-cultured across a membrane with human umbilical vein endothelial cells (HUVECs), do a subset of ADSCs show appropriate CD31 distribution along cell boundaries. Others have recently described that mesenchymal stem cells derive from perivascular cells including pericytes, which are known to wrap circumferentially around microvessels. We used ultrathin porous membranes to permit limited physical interactions between polarized HUVECs and ADSCs. In this arrangement, we found that the majority of ADSCs aligned perpendicular to the polarized HUVECs even though contact between the cell types was limited by pores less than 500 nm in diameter. Together, this ADSC pericyte behavior in combination with the ability to differentiate into ECs shows the potential versatility of ADSCs in the engineering of vascular networks.

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Acknowledgments

This work was supported, in part by the Gleason Family Foundation, RIT Seed Fund and the New York State Foundation for Science, Technology and Innovation (NYSTAR) & Center for Emerging and Innovative Sciences (CEIS). We thank Joshua Winans for assistance in SEM imaging of the membranes. We also thank James McGrath and the Nanomembrane Research Group (NRG) for helpful discussions throughout the study.

Conflict of interest

Andrea Mazzocchi and Alan Man declare that they have no conflict of interest. Jon-Paul S. DesOrmeaux is an employee of SiMPore and declares a potential conflict of interest. Thomas R. Gaborski is a co-founder and significant equity owner of SiMPore and declares a potential conflict of interest. SiMPore is a manufacturer of ultrathin silicon-based membranes and a co-sponsor of the NYSTAR Grant.

Ethical Standards

No human studies were carried out by the authors for this article. No animal studies were carried out by the authors for this article.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Rochester Institute of Technology, 160 Lomb Memorial Drive, Rochester, NY, 14623, USA

    Andrea R. Mazzocchi, Alan J. Man & Thomas R. Gaborski

  2. SiMPore, West Henrietta, NY, 14586, USA

    Jon-Paul S. DesOrmeaux

Authors
  1. Andrea R. Mazzocchi
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  2. Alan J. Man
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  4. Thomas R. Gaborski
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Corresponding author

Correspondence to Thomas R. Gaborski.

Additional information

Associate Editor Cynthia A. Reinhart-King oversaw the review of this article.

This paper is part of the 2014 Young Innovators Issue.

Thomas Gaborski completed a B.S. in Biological and Environmental Engineering from Cornell University in 2002 and a Ph.D. in Biomedical Engineering from the University of Rochester in 2008. As a graduate student, he was a university presidential fellowship recipient and also received a NIH Kirschstein NRSA predoctoral fellowship. His graduate work initially focused on neutrophil recruitment and the biophysics of adhesion molecule mobility and surface localization. It was during this work that Tom became involved with the life science applications of a novel class of ultrathin membranes leading to the co-founding of SiMPore in 2007. Tom initially served as head of life science application development at SiMPore and then as President from 2009 to 2012 during which time he helped lead the production, product development and membrane characterization teams. While at SiMPore, Tom was the principle investigator on several NIH small business innovative research grants. In 2012, Tom shifted his focus back towards academic research and joined the newly formed RIT Biomedical Engineering department. At RIT, his laboratory researches large-scale fabrication of ultrathin membranes and investigates cellular interactions on and across permeable substrates.

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Mazzocchi, A.R., Man, A.J., DesOrmeaux, JP.S. et al. Porous Membranes Promote Endothelial Differentiation of Adipose-Derived Stem Cells and Perivascular Interactions. Cel. Mol. Bioeng. 7, 369–378 (2014). https://doi.org/10.1007/s12195-014-0354-7

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