Authors Ofenbauer, A. ; Sebinger, D. ; Prewitz, M. ; Gruber, P. ; Werner, C.
Title Dewaxed ECM: A simple method for analyzing cell behaviour on decellularized extracellular matrices
Date 01.09.2015
Number 48041
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. Copyright © 2012 John Wiley & Sons, Ltd.
Publisher Journal of Tissue Engineering and Regenerative Medicine
Citation Journal of Tissue Engineering and Regenerative Medicine 9 (2015) 1046-1055
Tags decellularization extracellular matrix (ecm) kidney scaffold stem cells paraffin slide cell seeding differentiation

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