A 20 μl aliquot of expansion medium containing 2 × 10 chondrocytes (passage 2) was placed on the fibre side of each bilayered scaffold and cells were allowed to adhere for 2 h.
By direct mixing of those factors with biomaterials before the administration to the medium, the medium containing those mixture showed the chondrocyte growth of approximately a 25-fold increase by day 10.
In type II experiments, performed on rabbit chondrocytes, fresh, serum-free medium containing 0.1% BSA was added with or without IGF-1, IGFBP-3, and NBI-31772.
In type III experiments, performed on human OA chondrocytes, fresh, serum-free medium containing 0.1% BSA with or without IGF-1 or R3 IGF-1 was added to the cells and they were incubated for 24 hours.
Chondrocytes were grown in Ham F12 medium containing 10% FCS, 100 IU/ml penicillin, and 100 μg/ml streptomycin.
Following extraction, chondrocytes were cultured in DMEM culture medium containing 10% FBS, 2 mℳℒ-glutamine, 200 IU/ml penicillin, 200 μg/ml streptomycin and 40 IU/ml Nystatin.
During monolayer expansion chondrocytes were cultured in whole-cell culture medium containing 10% FCS.
Chondrocytes were washed three times with serum-starved medium (containing only 0.5% FCS) and further incubated for 30 min with the same medium before initiating treatment with LPS and/or inhibitors.
Chondrocytes were washed three times with serum-starved medium (containing only 3% FBS) and further incubated for 30 minutes with the same medium before initiation of treatment with TNF-β and/or inhibitors.
The chondrocyte-alginate constructs were then cultured in DMEM medium, containing 10% FBS and 1% penicillin-streptomycin, in a 37°C, 5% CO2 incubator for 3 to 4 weeks.
