目的:观察体外经传代培养去分化的关节软骨细胞,经关节康血清培养后生物学性状及功能的变化,探索使去分化软骨细胞再分化的手段,为软骨细胞组织工程种子细胞建立合适的体外培养方法。方法:无菌条件下取兔关节软骨组织,酶消化法分离软骨细胞,分成对照组及实验组,实验组常规培养3代后添加关节康血清,继续培养2周。观察对照各组细胞形态,细胞爬片后进行阿利新蓝染色及Ⅱ型胶原免疫组织化学检测,观察细胞表型变化。结果:兔关节软骨细胞体外培养3代后迅速去分化,增殖缓慢。添加关节康血清后去分化速度明显减缓,血清混合培养2周后,细胞外形恢复良好,阿利新蓝染色阳性及Ⅱ型胶原免疫组织化学检测阳性。结论:关节康血清与软骨细胞的混合培养,能延缓软骨细胞的去分化,并可以促进去分化后的软骨细胞再分化,分泌Ⅱ型胶原及糖蛋白,保持软骨细胞的形态和功能,有望用于在体外培养时防止软骨细胞的去分化及促进去分化软骨细胞的再分化。 OBJECTIVE: To observe the changes in biological characteristics and functions of dedifferentiated articular chondrocytes cultured in vitro through the subculture of kukangkang serum, and explore the means of redifferentiating the dedifferentiated chondrocytes in order to establish suitable in vitro cultured chondrocyte tissue engineering seed cells. Cultivation methods. METHODS: Rabbit articular cartilage tissue was taken aseptically and chondrocytes were isolated by enzyme digestion. The chondrocytes were divided into control group and experimental group. The experimental group was cultured for 3 generations followed by kang kang serum and cultured for 2 weeks. Observe the cell morphology of the control group. The cells were stained with alithio blue and type II collagen immunohistochemistry to observe the cell phenotype changes. RESULTS: Rabbit articular chondrocytes dedifferentiated rapidly after in vitro culture for 3 generations, and the proliferation was slow. The rate of dedifferentiation was significantly slowed after addition of kang kang sera. After 2 weeks of serum mixed culture, the cell morphology recovered well, alixin blue staining was positive and type II collagen immunohistochemistry was positive. Conclusion: The mixed culture of kang kang serum and chondrocyte can delay the dedifferentiation of chondrocytes, and can promote the redifferentiation of dedifferentiated chondrocytes, secrete type II collagen and glycoprotein, and maintain the morphology and function of chondrocytes. In vitro culture prevents the dedifferentiation of chondrocytes and promotes the differentiation of dedifferentiated chondrocytes.