pp 1-11

Part of the Methods in Molecular Biology book series

Decellularization of Large Tendon Specimens: Combination of Manually Performed Freeze-Thaw Cycles and Detergent Treatment

Protocol

Abstract

Reliable decellularization techniques applicable to tendon tissue play a critical role in the field of current tissue engineering. Particularly, an application as three-dimensional culture model for in vitro research and translational approaches to establish graft-based tendon repair as a routine clinical tool represent two main application fields for decellularized tendon scaffolds. Considering methodological issues of tendon decellularization, one of the major challenges lies in the preservation of the tendon-specific extracellular matrix (ECM) architecture to reflect natural tissue characteristic as best as possible. Concurrently, further requirements for high-quality decellularized biological tendon scaffolds include not only the reduction of resident cells, but also an ensured cytocompatibility.

To date, a large number and a wide variety of decellularization protocols for natural tendon tissue have already been investigated and usually, physical as well as chemical and/or enzyme-based treatments are used for the purpose of decellularization. However, to the best of our knowledge, there is a lack of evidence-based protocols for the processing of full-thickness large tendon samples, such as the equine flexor tendons.

Therefore, the here presented protocol describes a reliable procedure to decellularize equine superficial digital flexor tendons by using a combined treatment of physical decellularization in the form of repetitive freeze-thaw cycles, and of chemical decellularization with the non-ionic detergent Triton X-100. The decellularization effectiveness evaluated by reduction of cell and DNA content, the influence of decellularization on the morphology of the tendon extracellular matrix (ECM) as well as the cytocompatibility of the decellularized tendon scaffolds obtained have been investigated previously. Based on this previous study, the here present protocol is an effective procedure, particularly applicable for large tendon specimens.

Keywords:

Regenerative medicine Tissue engineering Decellularization Scaffold Freeze-thaw cycles Triton X-100 Detergent treatment Tendon Horse 

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Susanne Pauline Roth
    • 1
    • 2
  • Ina Erbe
    • 1
  • Janina Burk
    • 1
    • 3
  1. 1.Faculty of Veterinary Medicine, Equine Clinic and Hospital LeipzigUniversität LeipzigLeipzigGermany
  2. 2.Saxonian Incubator for Clinical TranslationUniversität LeipzigLeipzigGermany
  3. 3.Faculty of Veterinary Medicine, Institute of Veterinary PhysiologyUniversität LeipzigLeipzigGermany

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