pp 1-15 | Cite as

Decellularized Iliotibial Band Recolonized with Allogenic Homotopic Fibroblasts or Bone Marrow-Derived Mesenchymal Stromal Cells

  • Clemens Gögele
  • Silke Schwarz
  • Benjamin Ondruschka
  • Niels Hammer
  • Gundula Schulze-Tanzil
Protocol
Part of the Methods in Molecular Biology book series

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).

Keywords:

Acellularization Decellularization Explant culture Iliotibial band Ligamentocytes Mesenchymal stromal cells Recellularization 

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

Notes

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.

References

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Clemens Gögele
    • 1
  • Silke Schwarz
    • 1
  • Benjamin Ondruschka
    • 2
  • Niels Hammer
    • 3
  • Gundula Schulze-Tanzil
    • 1
  1. 1.Institute of AnatomyParacelsus Medical University, Salzburg and NurembergNurembergGermany
  2. 2.Institute of Legal MedicineUniversity of LeipzigLeipzigGermany
  3. 3.Department of Anatomy, Otago School of Biomedical SciencesUniversity of OtagoDunedinNew Zealand

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