Chondrocyte Isolation and Expansion

Harrison, Paul; Hopkins, Timothy; Hulme, Charlotte; McCarthy, Helen; Wright, Karina

Abstract

Chondrocyte isolation requires a combination of enzymatic and mechanical digestion of cartilaginous tissues in order to release the chondrocytes. Extracted primary chondrocytes will then adhere to standard tissue culture plastics, typically in small clusters, over a period of a few days in monolayer culture. Chondrocyte populations are expanded in a basal medium containing serum, supplemented with ascorbic acid, antibiotics, and sometimes antifungal agents and growth factors. Here we describe the standard research grade and good manufacturing practice (GMP) protocols used for the isolation and expansion of chondrocytes by the Oswestry/Keele University Orthopaedic Research (OsKOR) group and John Charnley GMP and MHRA licensed laboratory, both based at the RJAH Orthopaedic Hospital, Oswestry, UK.

• Online Publication Date: Nov 11, 2022
• Deposit Date: Jun 1, 2023
• Publisher: Springer
• Pages: 9-19
• Series Title: Methods in Molecular Biology
• Series ISSN: 1064-3745; 1940-6029
• Book Title: Cartilage Tissue Engineering
• DOI: https://doi.org/10.1007/978-1-0716-2839-3_2
• Keywords: Cartilage; Mechanical and enzymatic cartilage digestion; Chondrocytes; Tissue culture; Trypsinization; Autologous serum
• Additional Information: First Online: 11 November 2022