Optimisation of Chondrocyte Propagation from Human Articular Cartilage to Preserve the Chondrocyte characteristics
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Abstract
Introduction: Osteoarthritis (OA) is a major health burden and affects millions of people worldwide. This occurs when the cartilage degenerates over a period of time and OA commonly affects the joints in the hand, hips, and knees. Autologous chondrocyte implantation (ACI) and Matrix, matrix-associated autologous chondrocyte implantation (MACI) is a procedure used to treat isolated full-thickness articular cartilage defects of the knee. The cornerstone of these procedures is the ability to cultivate and grow a large number of chondrocytes for a successful outcome.
Materials and Methods: Cartilage specimens of patients undergoing total knee arthroplasty (TKA) were digested using standard collagenase treatment. The resulting cells isolated were evaluated for viability. The cells at equal seeding density were plated on different media for evaluation. Cells from each passage were counted, assessed for viability, immunophenotyping utilizing surface markers used for the characterization of chondrocytes and RT-PCR was performed to confirm expression of chondrocyte genes.
Results: Primary cells plated in DMEM F12 attained faster confluency, the addition of ITS supplement was not essential. Cells maintained their chondrocyte morphology and surface marker expression over passages and expressed genes for chondrocytes. Additional 5 samples were cultured on DMEM F12 without ITS and found to maintain chondrocyte characteristics.
Clinical Significance: Chondrocytes are crucial not only for the development of therapeutic approaches in cartilage repair but are necessary in cartilage tissue engineering to allow the development of functional cell models and novel scaffolds. As a result, an optimized, proven method for chondrocyte isolation must be developed and shared with other groups.
Conclusion: In this study, we describe a simple and affordable procedure of isolation and cultivation of human articular chondrocytes that demonstrated a high chondrogenic potential to the third passage, which is sufficient to grow cells for an ACI procedure.
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