Dmem nutrient mixture f 12 with glutamax dmem f12 glutamax

Human knee OA articular cartilage was obtained from the Department of Orthopedics at Gentofte University Hospital, Denmark, following international ethical guidelines for handling human samples and patient information (approved by the Danish National Committee on Biomedical Research Ethics, Department of Health, approval no. H D-2007-0084). All participants signed informed consent prior to enrolment in the study. The culture medium was composed of DMEM/Nutrient Mixture F-12 with GlutaMAX (DMEM/F12-GlutaMAX) with 1% Penicillin Streptomycin (PS) both obtained from Life Technologies. Phosphate buffered saline and human insulin-like growth factor-1 (IGF-1) were obtained from Sigma-Aldrich. FGF18 was obtained from Abcam. The sprifermin used was drug product formulated for intra-articular administration obtained from Merck KGaA together with the corresponding vehicle formulation. The vehicle formulation contained 7 mM Na2HPO4, 1 mM KH2PO4, 2.7 mM KCl (pH 7.3). Selection of doses was based on in vivo pharmacodynamic data published by Gigout et al. in 2017, where does between 100 and 1000 ng/mL were most effective^{11 (link)}. The doses were confirmed and narrowed to 300 to 900 ng/mL by studies in bovine cartilage explant published by Reker et al. in 2017^{12 (link)}.

The culture medium was composed of DMEM/Nutrient Mixture F-12 with GlutaMAX (DMEM/F12-GlutaMAX) with 1% Penicillin Streptomycin (PS) both obtained from Life Technologies. Phosphate buffered saline (PBS), human OSM and human insulin-like growth factor-I (IGF-I) were obtained from Sigma-Aldrich, and human TNF-α were from R&D systems. The sprifermin used was drug product formulated for intra-articular administration obtained from Merck-Serono together with the corresponding placebo formulation. The placebo formulation contained 7 mM Na₂HPO₄, 1.0 mM KH₂PO₄, 2.7 mM KCl (pH 7.3).