Dewaxed ECM: A simple method for analyzing cell behaviour on decellularized extracellular matrices

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Andreas Ofenbauer - , Leibniz Institute of Polymer Research Dresden (Author)
  • David Daniel Raphael Sebinger - , Leibniz Institute of Polymer Research Dresden (Author)
  • Marina Prewitz - , Leibniz Institute of Polymer Research Dresden (Author)
  • Petra Gruber - (Author)
  • Carsten Werner - , Chair of Biofunctional Polymer Materials, Leibniz Institute of Polymer Research Dresden (Author)

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 13days 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.

Details

Original languageEnglish
Pages (from-to)1046-1055
Number of pages10
JournalJournal of tissue engineering and regenerative medicine
Volume9
Issue number9
Publication statusPublished - 1 Sept 2015
Peer-reviewedYes

External IDs

PubMed 23172824
ORCID /0000-0003-0189-3448/work/161890451

Keywords

Keywords

  • Cell seeding, Decellularization, Differentiation, Extracellular matrix (ECM), Kidney, Paraffin slide, Scaffold, Stem cells