All chemicals were obtained from either Sigma-Aldrich (Copenhagen, Denmark) or Merck Millipore (Hellerup, Denmark) unless other stated. Recombinant human active ADAMTS-5 (cat#, 2198AD), human pro-MMP13 (cat#, 511-MM-010), human TNF-α (cat#, 210-TA) were purchased from R&D systems (Abingdon, UK). Dulbecco’s Modified Eagle Medium, Nutrient Mixture F-12 (DMEM: F12) was purchased from Life Technologies (Naerum, Denmark). AlamarBlue was from VWR international (Herlev, Copenhagen). Oncostatin M human (cat#, O9635) was from Sigma-Aldrich (Copenhagen, Denmark). GM6001 (cat#, M5939, Sigma-Aldrich) stock solution was dissolved in DMSO and then 1000x diluted in culture medium to a final concentration of 10μM. Proprotein Convertase (PC) inhibitor (cat#, 537076, Millipore) stock solution was prepared in DMSO and 1000x diluted in culture medium to a final concentration of 10μM. E64 (cat#, E3132, Sigma-Aldrich) was dissolved in sterile water to a final concentration of 10μM.
The streptavidin pre-coated ELISA plates were purchased from Roche (Mannheum, Germany). All ELISA plates were analyzed with the reader from Molecular Devices, SpectraMax M (CA, USA). Active ADAMTS-4, -5, MMP-9, -13, AGNxI and AGNxII lab scale kits were produced in NordicBioscience (Herlev, Denmark). The pre-clinical Cartilage-Laps ELISA was obtained from IDS Ltd. (Boldon, UK).
The streptavidin pre-coated ELISA plates were purchased from Roche (Mannheum, Germany). All ELISA plates were analyzed with the reader from Molecular Devices, SpectraMax M (CA, USA). Active ADAMTS-4, -5, MMP-9, -13, AGNxI and AGNxII lab scale kits were produced in NordicBioscience (Herlev, Denmark). The pre-clinical Cartilage-Laps ELISA was obtained from IDS Ltd. (Boldon, UK).