Abstract
Chondrocyte transplantation has been successfully tested and proposed as a clinical procedure aiming to repair articular cartilage defects. However, the isolation of chondrocytes and the optimization of the enzymatic digestion process, as well as their successful in vitro expansion, remain the main challenges in cartilage tissue engineering. In order to address these issues, we investigated the performance of recombinant collagenases in tissue dissociation assays with the aim of isolating chondrocytes from bovine nasal cartilage in order to establish the optimal enzyme blend to ensure the best outcomes of the overall procedure. We show, for the first time, that collagenase H activity alone is required for effective cartilage digestion, resulting in an improvement in the yield of viable cells. The extracted chondrocytes proved able to grow and activate differentiation/dedifferentiation programs, as assessed by morphological and gene expression analyses.
Highlights
Organ transplant therapy is currently hampered by the limited availability of compatible donors, and often by the onset of severe immune complications
The isolation of a large number chondrocytes from cartilage is a necessary and critical step for tissueThe repair/regeneration through
Adult cartilage tissue is composed of an abundant family of ECM proteins, among which collagens are the most representative, making their digestion an essential step for the proper purification of cells
Summary
Organ transplant therapy is currently hampered by the limited availability of compatible donors, and often by the onset of severe immune complications. Chondrocytes have been shown to be among the most versatile cell types due to their renewal/differentiation ability, which makes them suitable for various applications [2] In this context, autologous chondrocyte implantation (ACT) is considered the gold standard for cartilage repair [3]; recent preclinical studies have provided promising evidence for the effectiveness of cell-based regeneration strategies [4,5]. Adult Nasal Chondrocytes (NC) are considered worthy of attention due to their unique features [7] They derive from the neural crest, and have been recently shown to be able to respond and adapt to heterotopic transplantation sites.
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