Use of enzymatically isolated chondrocytes for short term metabolic studies

Chondrocytes isolated from calf articular cartilage by pretreatment with various enzymes prior to collagenase digestion were examined to determine the effect of enzymatic exposure on cell viability and to establish optimal conditions whereby freshly isolated cells could be used for short term metabolic studies. Pre-collagenase exposure to proteolytic enzymes for short intervals has no effect on cell viability. Trypsin pre-treatment appeared to increase the efficiency of cell isolation. Proteoglycan and RNA synthetic activity was influenced by the conditions of incubation (i.e., cell suspension vs. cell pellet: with or without a 24 hour "rest" period; and differential pre-collagenase enzyme exposure). The data suggest that the experimental conditions of incubation and the enzymes used in isolation of the cells, materially alter the freshly isolated chondrocyte's "pattern" of metabolic activity in short term culture studies.