Abstract:

BACKGROUND: Recent years, most studies have attempted to repair defected articular cartilage using isolated articular cartilage cells. However, it is difficult to obtain purified articular cartilage cells that have biological activity.
OBJECTIVE: To harvest articular cartilage cells using combined digestion of trypsin and type Ⅱ collagen.
METHODS: Articular cartilages were obtained from the surface of normal femoral head of SD rats, digested by trypsin (0.25%) and collagease Ⅱ (0.2%). After observing numerous articular cartilage cells free under a phase contrast microscope, the big pieces of articular cartilage was abandoned, followed by centrifugation and washing with phosphate buffered saline for 2 times. And then, the mixture was cultured in cartilage cells culture medium. Toluidine blue and hematoxylin-eosin staining were used to identify the articular cartilage cells.
RESULTS AND CONCLUSION: With seriously controlling the concentrate of enzyme and the time of digestion, through enzymolysis of articular cartilage cells by trypsin (0.25%) and type Ⅱ collagease (0.2%). Articular cartilage cells were obtained by isolating and cultivating from femoral shaft of SD rats, which was identified with toluidine blue and hematoxylin-eosin staining.