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Dewaxed ECM: A simple method for analyzing cell behaviour on decellularized extracellular matrices
Author(s) -
Ofenbauer Andreas,
Sebinger David Daniel Raphael,
Prewitz Marina,
Gruber Petra,
Werner Carsten
Publication year - 2015
Publication title -
journal of tissue engineering and regenerative medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.835
H-Index - 72
eISSN - 1932-7005
pISSN - 1932-6254
DOI - 10.1002/term.1658
Subject(s) - decellularization , extracellular matrix , tissue engineering , biomedical engineering , chemistry , stem cell , embryonic stem cell , regenerative medicine , cell , microbiology and biotechnology , biology , gene , medicine , biochemistry
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.