Julius-Maximillians University, Würzburg, Germany

6 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.