目的 :研究关节软骨细胞的原代分离及体外培养 ,了解表皮细胞生长因子 ( EGF)对其生长和去分化趋势的影响。方法 :( 1)用自制消化瓶分离原代软骨细胞并培养、传代 ;( 2 )锥虫蓝染色测定细胞存活率 ,测定克隆率 ;( 3 )比较 EGF对软骨细胞增殖以及去分化趋势的影响。 结果 :( 1)自制消化瓶能在 3 h内将软骨碎块消化完毕 ;( 2 )细胞存活率为 88.4% ,克隆率为2 9 .4% ;( 3 ) EGF显著促进软骨细胞增殖 ( P<0 .0 5 ) ;( 4 )软骨细胞经过 8～ 10次传代以后出现去分化趋势 ,与培养液中是否含EGF无关。 结论 :自制消化瓶能提高消化软骨的效率。 EGF能显著促进软骨细胞增殖 ,但不能阻止软骨细胞的去分化趋势 OBJECTIVE: To study the primary isolation and culture of articular chondrocytes in vitro and to investigate the effect of epidermal growth factor (EGF) on their growth and dedifferentiation. Methods: (1) Primary chondrocytes were isolated and cultured with self-made digestion vials and passaged; (2) Cell viability was assayed by trypan blue staining to determine the clonality; (3) Effects of EGF on chondrocyte proliferation and dedifferentiation . (2) Cell survival rate was 88.4%, the cloning rate was 29.4%; (3) EGF significantly promoted the proliferation of chondrocytes (P <0 .0 5). (4) Chondrocytes tended to dedifferentiate after 8 to 10 passages, regardless of whether they contained EGF or not. Conclusion: Self-made digestion bottles can improve the efficiency of digestion cartilage. EGF can significantly promote chondrocyte proliferation, but can not prevent the dedifferentiation trend of chondrocytes