Biomolecules Based Microarray for Screening Human Endothelial Cells Behavior
Endothelial Progenitor Cell (EPC) based therapies continue to be of interest to treat ischemic events based on their proven role to promote blood vessel formation and thus tissue re-vascularisation. Current strategies for the production of clinical-grade EPCs requires the in vitro isolation of EPCs from peripheral blood followed by cell expansion to provide sufficient quantities EPCs for cell therapy. This study aims to examine the use of different biomolecules to significantly improve the current strategy of EPC capture and expansion on collagen type I (Col I). In this study, four different biomolecules were immobilised on a surface and then investigated for their capacity to support EPC capture and proliferation. First, a cell microarray platform was fabricated by coating a glass surface with epoxy functional allyl glycidyl ether plasma polymer (AGEpp) to mediate biomolecule binding. The four candidate biomolecules tested were Col I, collagen type II (Col II), collagen type IV (Col IV) and vascular endothelial growth factor A (VEGF-A), which were arrayed on the epoxy-functionalised surface using a non-contact printer. The surrounding area between the printed biomolecules was passivated with polyethylene glycol-bisamine (A-PEG) to prevent non-specific cell attachment. EPCs were seeded onto the microarray platform and cell numbers quantified after 1 h (to determine capture) and 72 h (to determine proliferation). All of the extracellular matrix (ECM) biomolecules printed demonstrated an ability to capture EPCs within 1 h of cell seeding with Col II exhibiting the highest level of attachment when compared to the other biomolecules. Interestingly, Col IV exhibited the highest increase in EPC expansion after 72 h when compared to Col I, Col II and VEGF-A. These results provide information for significant improvement in the capture and expansion of human EPC for further application.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1128253Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1256
 Murray, C.J.L. and A.D. Lopez Measuring the Global Burden of Disease. New England Journal of Medicine, 2013. 369(5): p. 448-457.
 Sen, S., et al., Endothelial progenitor cells: novel biomarker and promising cell therapy for cardiovascular disease. Clinical science, 2011. 120(7): p. 263-283.
 Cho, H.-J., et al., Mobilized Endothelial Progenitor Cells by Granulocyte-Macrophage Colony-Stimulating Factor Accelerate Reendothelialization and Reduce Vascular Inflammation After Intravascular Radiation. Circulation, 2003. 108(23): p. 2918-2925.
 Colombo, E., et al., Comparison of fibronectin and collagen in supporting the isolation and expansion of endothelial progenitor cells from human adult peripheral blood. PloS one, 2013. 8(6): p. e66734.
 Yang, J., et al., CD34+ cells represent highly functional endothelial progenitor cells in murine bone marrow. PloS one, 2011. 6(5): p. e20219.
 Dimmeler, S., J. Burchfield, and A.M. Zeiher, Cell-based therapy of myocardial infarction. Arteriosclerosis, thrombosis, and vascular biology, 2008. 28(2): p. 208-216.
 Hu, Y., et al., Endothelial Replacement and Angiogenesis in Arteriosclerotic Lesions of Allografts Are Contributed by Circulating Progenitor Cells. Circulation, 2003. 108(25): p. 3122-3127.
 Wassmann, S., et al., Improvement of endothelial function by systemic transfusion of vascular progenitor cells. Circulation Research, 2006. 99(8): p. E74-E83.
 Sukmawati, D. and R. Tanaka, Introduction to next generation of endothelial progenitor cell therapy: a promise in vascular medicine. Am J Transl Res, 2015. 7(3): p. 411-421.
 Kamata, S., et al., Improvement of Cardiac Stem Cell Sheet Therapy for Chronic Ischemic Injury by Adding Endothelial Progenitor Cell Transplantation: Analysis of Layer-Specific Regional Cardiac Function. Cell transplantation, 2014. 23(10): p. 1305-1319.
 Atesok, K., R. Li, and E. Schemitsch, Endothelial Progenitor Cells: A Novel Cell‐based Therapy in Orthopaedic Surgery. Journal of the American Academy of Orthopaedic Surgeons, 2012. 20(10): p. 672-674.
 Kaneko, Y., et al., Cell therapy for stroke: emphasis on optimizing safety and efficacy profile of endothelial progenitor cells. Current pharmaceutical design, 2012. 18(25): p. 3731-3734.
 Assmus, B., et al., Transplantation of progenitor cells and regeneration enhancement in acute myocardial infarction (TOPCARE-AMI). Circulation, 2002. 106(24): p. 3009-3017.
 Anderson, D.G., S. Levenberg, and R. Langer, Nanoliter-scale synthesis of arrayed biomaterials and application to human embryonic stem cells. Nature Biotechnology, 2004. 22(7): p. 863-866.
 Titmarsh, D.M., et al., Arrayed cellular environments for stem cells and regenerative medicine. Biotechnology journal, 2013. 8(2): p. 167-179.
 Peters, A., D.M. Brey, and J.A. Burdick, High-throughput and combinatorial technologies for tissue engineering applications. Tissue Engineering Part B: Reviews, 2009. 15(3): p. 225-239.
 Anglin, E., et al., Cell microarrays for the screening of factors that allow the enrichment of bovine testicular cells. Cytometry Part A, 2010. 77(9): p. 881-889.
 Hook, A.L., H. Thissen, and N.H. Voelcker, Surface manipulation of biomolecules for cell microarray applications. TRENDS in Biotechnology, 2006. 24(10): p. 471-477.
 Nakajima, M., et al., Combinatorial protein display for the cell-based screening of biomaterials that direct neural stem cell differentiation. Biomaterials, 2007. 28(6): p. 1048-1060.
 Martin-Ramirez, J., et al., Establishment of outgrowth endothelial cells from peripheral blood. Nat. Protocols, 2012. 7(9): p. 1709-1715.
 Dalilottojari, A., et al., Porous silicon based cell microarrays: optimizing human endothelial cell-material surface interactions and bioactive release. Biomacromolecules, 2016.
 Delalat, B., et al., A Combinatorial Protein Microarray for Probing Materials Interaction with Pancreatic Islet Cell Populations. Microarrays, 2016. 5(3): p. 21.
 Browning, A., H. Dua, and W. Amoaku, The effects of growth factors on the proliferation and in vitro angiogenesis of human macular inner choroidal endothelial cells. British Journal of Ophthalmology, 2008. 92(7): p. 1003-1008.
 Siavashi, V., et al., ECM‐Dependence of Endothelial Progenitor Cell Features. Journal of cellular biochemistry, 2016.
 Yang, N., et al., The characteristics of endothelial progenitor cells derived from mononuclear cells of rat bone marrow in different culture conditions. Cytotechnology, 2011. 63(3): p. 217-226.
 Liu, X., et al., Regulatory effects of soluble growth factors on choriocapillaris endothelial growth and survival. Ophthalmic Research, 1998. 30(5): p. 302-313.
 Rasi Ghaemi, S., et al., Surface engineering for long-term culturing of mesenchymal stem cell microarrays. Biomacromolecules, 2013. 14(8): p. 2675-2683.
 Ataollahi, F., et al., Endothelial cell responses in terms of adhesion, proliferation, and morphology to stiffness of polydimethylsiloxane elastomer substrates. Journal of Biomedical Materials Research Part A, 2015. 103(7): p. 2203-2213.