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Universitätsklinikum Würzburg

7 ARTICLES PUBLISHED IN JoVE

Bioengineering

Non-contact, Label-free Monitoring of Cells and Extracellular Matrix using Raman Spectroscopy

Miriam Votteler^1,2, Daniel A. Carvajal Berrio², Marieke Pudlas^2,3, Heike Walles^2,4, Katja Schenke-Layland^1,2

¹Department of Thoracic and Cardiovascular Surgery and Inter-University Centre for Medical Technology Stuttgart-Tübingen (IZST), Eberhard Karls University, Tübingen, ²Department of Cell and Tissue Engineering, Fraunhofer Institute of Interfacial Engineering and Biotechnology (IGB) Stuttgart, Germany, ³Department for Medical Interfacial Engineering (IGVT), University of Stuttgart, Germany, ⁴Institute of Tissue Engineering and Regenerative Medicine, Julius-Maximillians University, Würzburg, Germany

Raman spectroscopy is a suitable technique for the non-contact, label-free analysis of living cells, tissue-engineered constructs and native tissues. Source-specific spectral fingerprints can be generated and analyzed using multivariate analysis.

Bioengineering

Tissue Engineering of a Human 3D in vitro Tumor Test System

Corinna Moll^*1, Jenny Reboredo^*1, Thomas Schwarz¹, Antje Appelt¹, Sebastian Schürlein¹, Heike Walles¹, Sarah Nietzer¹

¹Department of Tissue Engineering and Regenerative Medicine, University Hospital Würzburg

Methods to create human 3D tumor tissues as test systems are described. These technologies are based on a decellularized Biological Vascularized Scaffold (BioVaSc), primary human cells and a tumor cell line, which can be cultured under static as well as under dynamic conditions in a flow bioreactor.

Bioengineering

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

Angela Rossi^*1, Antje Appelt-Menzel^*1, Szymon Kurdyn¹, Heike Walles^1,2, Florian Groeber²

¹Department for Tissue Engineering and Regenerative Medicine, University Hospital Würzburg, ²Translational Center Würzburg, Regenerative Therapies in Oncology and Musculoskelettal Disease, Würzburg Branch of the Fraunhofer-Institute Interfacial Engineering and Biotechnology, IGB

The goal of this protocol is to build up a three-dimensional full thickness skin equivalent, which resembles natural skin. With a specifically constructed automated wounding device, precise and reproducible wounds can be generated under maintenance of sterility.

Bioengineering

A Combined 3D Tissue Engineered In Vitro/In Silico Lung Tumor Model for Predicting Drug Effectiveness in Specific Mutational Backgrounds

Claudia Göttlich^*1, Lena C. Müller^*1, Meik Kunz^*3, Franziska Schmitt¹, Heike Walles^1,4, Thorsten Walles², Thomas Dandekar³, Gudrun Dandekar^1,4, Sarah L. Nietzer¹

¹Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Wuerzburg, ²Department of Cardiothoracic Surgery, University Hospital Wuerzburg, ³Department of Bioinformatics, University Wuerzburg, ⁴Translational Center Wuerzburg, Fraunhofer Institute Interfacial Engineering and Biotechnology IGB

We present a three-dimensional (3D) lung cancer model based on a biological collagen scaffold to study sensitivity towards non-small-cell-lung-cancer-(NSCLC)-targeted therapies. We demonstrate different read-out techniques to determine the proliferation index, apoptosis and epithelial-mesenchymal transition (EMT) status. Collected data are integrated into an in silico model for prediction of drug sensitivity.

Bioengineering

Site-Directed Immobilization of Bone Morphogenetic Protein 2 to Solid Surfaces by Click Chemistry

Claudia Siverino¹, Barbara Tabisz², Tessa Lühmann³, Lorenz Meinel³, Thomas Müller⁴, Heike Walles^1,2, Joachim Nickel^1,2

¹Fraunhofer-Institut für Grenzflächen- und Bioverfahrenstechnik (IGB), Translationszentrum Würzburg 'Regenerative Therapien für Krebs- und Muskuloskelettale Erkrankung', Institutsteil Würzburg, ²Lehrstuhl für Tissue Engineering und Regenerative Medizin, Universitätsklinikum Würzburg, ³Lehrstuhl für Pharmazeutische Technologie und Biopharmazie, Universität Würzburg, ⁴Lehrstuhl für molekulare Pflanzenphysiologie und Biophysik, Julius-von-Sachs Institut für Biowissenschaften, Universität Würzburg

Biomaterials doped with Bone Morphogenetic Protein 2 (BMP2) have been used as a new therapeutic strategy to heal non-union bone fractures. To overcome side effects resulting from an uncontrollable release of the factor, we propose a new strategy to site-directly immobilize the factor, thus creating materials with improved osteogenic capabilities.

Bioengineering

Recombinant Collagen I Peptide Microcarriers for Cell Expansion and Their Potential Use As Cell Delivery System in a Bioreactor Model

Melva Suarez Muñoz¹, Davide Confalonieri¹, Heike Walles^1,2, Elisabeth M. W. M. van Dongen³, Gudrun Dandekar^1,2

¹Department Tissue Engineering and Regenerative Medicine, University Hospital Wuerzburg, ²Translational Center Regenerative Therapies (TLC-RT), Fraunhofer Institute for Silicate Research ISC, ³Fujifilm Manufacturing Europe B.V.

We propose a cell expansion protocol on macroporous microcarriers and their use as delivery system in a perfusion bioreactor to seed a decellularized tissue matrix. We also include different techniques to determine cell proliferation and viability of cells cultured on microcarriers. Furthermore, we demonstrate functionality of cells after bioreactor cultures.

Immunology and Infection

Engineering Chimeric Antigen Receptor-Natural Killer Cells Targeting Fungal Infections Using the Non-viral Sleeping Beauty Transposon System

Maximilian Bauser¹, Hermann Einsele¹, Jürgen Löffler¹, Michael Hudecek^1,2, Michelle Seif¹

¹Medizinische Klinik und Poliklinik II und Lehrstuhl für Zelluläre Immuntherapie, Medizinische Klinik und Poliklinik II, Universitätsklinikum Würzburg, ²Fraunhofer-Institut für Zelltherapie und Immunologie IZI, Außenstelle Würzburg Zelluläre Immuntherapie

We outline a transfection protocol for producing chimeric antigen receptor-natural killer (CAR-NK) cells targeting fungal pathogens using the non-viral Sleeping Beauty transposon system. To assess antigen-specific activation, we co-cultured the engineered cells with Aspergillus fumigatus germ tubes and measured IFN-γ secretion.