title: Dewaxed ECM: A simple method for analyzing cell behaviour on decellularized extracellular matrices.
abstract: Decellularization techniques have been used on a wide variety of tissues to create cell-seedable scaffolds for tissue engineering. Finding a suitable decellularization protocol for a certain type of tissue can be laborious, especially when organ perfusion devices are needed. In this study, we report a quick and simple method for comparing decellularization protocols combining the use of paraffin slices and two-dimensional cell cultures. We developed three decellularization protocols for adult murine kidney that yielded decellularized extracellular matrices (ECMs) with varying histological properties. The resulting paraffin-embedded ECM slices were deparaffinized and reseeded with murine embryonic stem cells (mESCs). We analyzed cell attachment four days post seeding via determination of cell numbers, and used quantitative Real-Time PCR 13 days post seeding to measure gene expression levels of two genes associated with renal development, Pax2 and Pou3f3. The three decellularization protocols produced kidney-matrices that showed clearly distinguishable results. We demonstrated that formerly paraffin-embedded decellularized ECMs can effectively influence differentiation of stem cells. This method can be used to identify optimal decellularization protocols for recellularization of three-dimensional tissue-scaffolds with embryonic stem cells and other tissue-specific cell types.
authors.affiliation: Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials, Hohe Str. 6, 01069, Dresden, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Ofenbauer
authors.affiliation: Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials, Hohe Str. 6, 01069, Dresden, Germany.
authors.forename: David Daniel Raphael
authors.initials: DD
authors.lastname: Sebinger
authors.affiliation: Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials, Hohe Str. 6, 01069, Dresden, Germany.
authors.forename: Marina
authors.initials: M
authors.lastname: Prewitz
authors.affiliation: Private contribution, Vienna.
authors.forename: Petra
authors.initials: P
authors.lastname: Gruber
authors.affiliation: Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials, Hohe Str. 6, 01069, Dresden, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Werner
mesh_terms: D000818:Animals; D053595:Embryonic Stem Cells / Q000166:cytology; D005109:Extracellular Matrix; D005260:Female; D066298:In Vitro Techniques; D007668:Kidney / Q000166:cytology; D051379:Mice; D016612:Paraffin Embedding; D060888:Real-Time Polymerase Chain Reaction; D054457:Tissue Scaffolds