Optimisation of isolation protocol of local bovine articular chondrocytes

The crosslinks and microstructure of cartilage are poorly understood and there are many different techniques to isolate chondrocytes from their extracellular matrix. The purpose of this study is to obtain an optimised protocol to harvest the highest number of viable cells from a particular species of cow, Bos indicus, which has not been previously reported. Therefore, this would allow the preparation of a sufficient quantity of cells for high seeding density, which in turn promotes the expression of normal chondrocytic phenotype and facilitates cartilage repair. Cartilage explants were digested in 1-hour pronase, followed by digestion in collagenase type-Ia, type-II or type-XI for duration of 14 to 16 hours. The concentration of the collagenase was either 100 or 200 U/ml. The results demonstrate that maximum cell yield was achieved from a two-step digestion with 1-hour digestion with pronase and 16-hours in collagenase type-II, without jeopardizing cell viability.