Advancement of clickECM derived from human differentiated stem cells for the use as tissue-specific surface coatings
Frontiers in Bioengineering and Biotechnology(2016)
Abstract
Event Abstract Back to Event Advancement of clickECM derived from human differentiated stem cells for the use as tissue-specific surface coatings Silke Keller1, Sybil M. Ruff2, Gabriele Vacun1, Günter E. Tovar1, 2, Monika Bach1, 2 and Petra J. Kluger1, 3 1 Fraunhofer Institute for Interfacial Engineering and Biotechnology, Cell and Tissue Engineering, Germany 2 University of Stuttgart, Institute of Interfacial Process Engineering and Plasma Technology, Germany 3 Reutlingen University, School of Applied Chemistry, Germany Human Mesenchymal Stem Cells (hMSCs) are adult multipotent stem cells and capable of differentiating into different connective tissue cells of the osteogenic, the adipogenic, or the chondrogenic lineage. These cells synthesize and secret proteins, proteoglycans, and glycosaminoglycans into the intercellular space to build a tissue-specific extracellular matrix (ECM). This ECM can be considered as the natural environment of the cells. The distinct tissue-specific properties of these matrices and their functions make the ECM exceedingly interesting as a biocompatible and bioactive coating for biomaterials[1],[2]. However, a major limitation of conventional ECM coatings is the lack of mechanical stability since the adhesion of the biomolecules is only based on physisorption[3]. Therefore, we developed a "clickable" ECM (clickECM) derived from dermal fibroblasts which can be covalently immobilized on artificial surfaces (Fig 1). Here we demonstrate that this technology is transferable to differentiated hMSC to generate a clickECM with tissue-specific properties for the use in tissue engineering and regenerative medicine. To generate tissue-specific clickECMs we treated human mesenchymal stem cells over a period of 7-21 days with an azide-modified monosaccharide (Ac4GalNAz, metabolic oligosaccharide engineering, MOE) either in stem cell media or in differentiation media (adipogenic, osteogenic, or chondrogenic). Next, cells were removed by lysis and the clickECMs were purified by washing with ultrapure water. Immunofluorescence staining of the typical ECM proteins collagen I, collagen IV, and fibronectin was used to analyze the protein expression in clickECMs depending on the differentiation status of the cells. Next, the tissue-specific properties were characterized using quantitative assays such as cell proliferation assays (DNA quantification), or the alkaline phosphatase (ALP) level as a marker for osteogenesis. We could show that MOE is a useful tool to introduce click groups into the ECM of MSC cell cultures. The click groups were ubiquitously distributed in the glycans of the isolated ECMs of both undifferentiated as well as differentiated MSCs. However, the protein level of collagen I and collagen IV varied depending on the differentiation status of the cells, supporting the hypothesis that cell differentiation results in a tissue-specific composition of the ECM. We could show that this has an influence on the biological activity of the ECM. The bone-like clickECM from osteogenically differentiated hMSCs for instance increases the ALP level of osteoblasts (C2C12 cell line) after five days of cell culture (Fig 2). In future, this clickECM could be covalently immobilized on hip prosthesis using copper-free click chemistry[4] to support the osseointegration. In contrast, a fatty tissue-like clickECM from adipogenically differentiated MSCs might improve the integration of breast implants, whereas the clickECM derived from undifferentiated hMSCs might be used as a coating which mimics the stem cell niche. In future these tissue-specific coatings could be used to facilitate the integration of medical implants into the patient’s body by offering the cells the optimal, tissue-specific conditions for growth and proliferation at the tissue-implant-interface. References:[1] Fitzpatrick, L. E. & McDevitt, T. C. Cell-derived matrices for tissue engineering and regenerative medicine applications. Biomater. Sci. 3 (2015).[2] Zhang, Y. Y. et al. Tissue-specific extracellular matrix coatings for the promotion of cell proliferation and maintenance of cell phenotype. Biomaterials 30, 4021-4028 (2009).[3] Goodman, S. B., Yao, Z. Y., Keeney, M. & Yang, F. The future of biologic coatings for orthopaedic implants. Biomaterials 34, 3174-3183 (2013).[4] Kolb, H. C., Finn, M. G. & Sharpless, K. B. Click chemistry: Diverse chemical function from a few good reactions. Angew. Chem. Int. Ed. Engl. 40, 2004-2021 (2001). Keywords: Extracellular Matrix, stem cell, Surface modification, matrix-cell interaction Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Bioconjugates Citation: Keller S, Ruff SM, Vacun G, Tovar GE, Bach M and Kluger PJ (2016). Advancement of clickECM derived from human differentiated stem cells for the use as tissue-specific surface coatings. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02440 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Silke Keller Sybil M Ruff Gabriele Vacun Günter E Tovar Monika Bach Petra J Kluger Google Silke Keller Sybil M Ruff Gabriele Vacun Günter E Tovar Monika Bach Petra J Kluger Google Scholar Silke Keller Sybil M Ruff Gabriele Vacun Günter E Tovar Monika Bach Petra J Kluger PubMed Silke Keller Sybil M Ruff Gabriele Vacun Günter E Tovar Monika Bach Petra J Kluger Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.
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Key words
Tissue-Specific Coatings,Tissue Engineering,Extracellular Matrix,Cardiac Tissue Engineering,Mesenchymal Stem Cells
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