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Decellularisation Process to Produce Graft for Transplantation; Repopulation Capacity of Human MSCS

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Shortage of graft for tissue or organ replacement until currently is still an issue worldwide. There are many source options for the graft transplantation, such as autograft, allograft and synthetics, however none of them are ideal. Currently, a technique to remove cells from an allogeneic or xenogeneic tissue while retaining matrix histoarchitecture is promising to generate an acellular biological graft, which possesses regeneration capability comparable to the autograft. In this study, the capability of the decellularized graft to allow cellular repopulation to initiate regeneration process was investigated. porcine patellar tendon (PPT) was used as the source of the graft. They were decellularised (n = 20) using chemical, mechanical, physical and enzymatic processes. The decellularised tendon scaffolds were analysed for their acellularity, ECM structure and components, quantity of DNA remains, sterility and biocompatibility. Human mesenchymal stem cells (human MSCs) were analysed for their mesenchymal properties. Seeding techniques were investigated; using seeding ring (n = 3) and direct method (n = 3). Both conditions had the target seeding density of 105 cells ยท cm−2. The capability of the decellularised scaffold to facilitate cellular attachment was measured using viability assay; qualitatively using Live/Dead staining (Life technology) and quantitatively using ATP counts (Perkin Elmer). all decellularised tendon scaffolds had fulfilled the criteria for an acellular graft and human MSCs possessed mesenchymal properties. Both seeding methods showed cellular attachment. Viability assay of cells seeded using seeding rings was significantly lower compared to the direct method (p < 0.05). acellular tendon scaffold using current decellularisation protocol had generated tendon scaffold that allowed cellular attachment.
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Keywords: Biological Graft; Decellularization; Scaffold; Seeding Technique; Viability Assay

Document Type: Research Article

Affiliations: Institute of Medical and Biological Engineering, University of Leeds, LS1 9JT, UK

Publication date: September 1, 2018

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