RJAH Orthopaedic Hospital NHS Foundation Trust, Oswestry, Shropshire, UK.
School of Pharmacy and Bioengineering, Keele University, Staffordshire, UK.
Methods Mol Biol. 2023;2598:9-19. doi: 10.1007/978-1-0716-2839-3_2.
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.
软骨细胞的分离需要结合酶解和机械消化软骨组织，以释放软骨细胞。提取的原代软骨细胞随后会在单层培养中，经过几天的时间，以小簇的形式附着在标准的组织培养塑料上。软骨细胞群体在含有血清的基础培养基中扩增，补充抗坏血酸、抗生素，有时还添加抗真菌药物和生长因子。这里我们描述了由 Oswestry/Keele 大学骨科研究（OsKOR）组和 John Charnley GMP 和 MHRA 许可实验室使用的标准研究级和良好生产规范（GMP）方案，这两个实验室都位于英国 Oswestry 的 RJAH 骨科医院。
