Abstract:

BACKGROUND: To establish an economic, efficient, practical isolation and culture system of chondrocytes has important significance to cartilage in vitro experiments.
OBJECTIVE: To investigate and improve the culture method of rat articular chondrocytes.
METHODS: Rat cartilage of bilateral hip and knee was resected from one week old male rats under aseptic condition. Chondrocytes were isolated by typeⅡ collagenase enzyme digestion method. The cells were cultured and passaged, then identified.
RESULTS AND CONCLUSION: Inverted phase contrast microscope observation showed that the primary cultured chondrocytes adherented at the 12th hour after cultivation. The monolayer formation occurred on the 3rd day after cultivation. And cells were ready to be passaged on the 4th day after cultivation. After passaged for six generations, some cells represented a spindle-like appearance. In the 7th generation, most cells turned into a long and irregular appearance, and cell proliferation capacity diminished. Toluidine blue staining showed that the nucleis of cultured chondrocytes were metachromatic. Immunofluorescent staining showed that cultured chondrocytes had a positive expression of collagen type Ⅱ. These findings illustrate that a large number of purified rat chondrocytes can be gained by this method in a short time.