Dewaxed ECM: A simple method for analyzing cell behaviour on decellularized extracellular matrices
Publikation: Beitrag in Fachzeitschrift › Forschungsartikel › Beigetragen › Begutachtung
Beitragende
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
Originalsprache | Englisch |
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Seiten (von - bis) | 1046-1055 |
Seitenumfang | 10 |
Fachzeitschrift | Journal of tissue engineering and regenerative medicine |
Jahrgang | 9 |
Ausgabenummer | 9 |
Publikationsstatus | Veröffentlicht - 1 Sept. 2015 |
Peer-Review-Status | Ja |
Externe IDs
PubMed | 23172824 |
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ORCID | /0000-0003-0189-3448/work/161890451 |
Schlagworte
ASJC Scopus Sachgebiete
Schlagwörter
- Cell seeding, Decellularization, Differentiation, Extracellular matrix (ECM), Kidney, Paraffin slide, Scaffold, Stem cells