Abstract
Decellularized scaffolds present promising biomimetic approaches in various fields of tissue engineering. Different tissues have been selected for decellularization, among them extracellular matrix (ECM)-rich tissues such as tendons, ligaments and cartilage. The dense ECM of ligaments is particularly challenging to achieve a completely non-immunogenic ECM void of any cells. Here, the methods for decellularization adapted to ligamentous tissue of the iliotibial band (ITB) are presented along with cell isolation and several recolonization techniques using allogenic ITB-derived fibroblasts or mesenchymal stromal cells (MSCs).
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Abbreviations
- AB:
-
Alcian blue
- ACL:
-
Anterior cruciate ligament
- dest:
-
Aqua destillata
- BSA:
-
Bovine serum albumin
- DAPI:
-
4′,6-Diamidino-2-phenylindol
- DMSO:
-
Dimethylsulfoxide
- DMEM:
-
Dulbecco’s Minimum Essential Medium
- DP:
-
Decellularization process
- ECM:
-
Extracellular matrix
- EDTA:
-
Ethylenediaminetetraacetic acid
- FBS:
-
Foetal bovine serum
- FDA:
-
Fluorescein diacetate
- HE:
-
Hematoxylin and eosin
- ITB:
-
Iliotibial band
- MSC:
-
Mesenchymal stromal cells
- MTS:
-
3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium
- PBS:
-
Phosphate buffered saline
- PET:
-
Polyethylene terephthalate
- PI:
-
Propidium iodide
- rpm:
-
Rounds per minute
- RPMI:
-
Roswell Park Memorial Institute medium
- RT:
-
Room temperature
- SDS:
-
Sodium dodecyl sulphate
- 3D:
-
Three-dimensional
- TBS:
-
Tris buffered saline
- TRIS:
-
Tris-(hydroxymethyl)-aminomethane
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Acknowledgement
We would like to thank Tobias Schneider for his support. The authors acknowledge the financial support from Manfred Roth foundation. We would like to thank the University of Salzburg and the Austrian Agency for International Cooperation in Education and Research (OeAD-GmbH) for the scholarship and financial support of Mr. Clemens Gögele.
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© 2017 Springer Science+Business Media New York
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Gögele, C., Schwarz, S., Ondruschka, B., Hammer, N., Schulze-Tanzil, G. (2017). Decellularized Iliotibial Band Recolonized with Allogenic Homotopic Fibroblasts or Bone Marrow-Derived Mesenchymal Stromal Cells. In: Turksen, K. (eds) Decellularized Scaffolds and Organogenesis. Methods in Molecular Biology, vol 1577. Humana Press, New York, NY. https://doi.org/10.1007/7651_2017_30
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DOI: https://doi.org/10.1007/7651_2017_30
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7655-3
Online ISBN: 978-1-4939-7656-0
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