关节软骨细胞的分离、培养及表皮细胞生长因子对其影响

Isolation and culture of articular chondrocytes and the effect of epidermal growth factor on it

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 。

Objective: To study the isolation and culture of chondrocyte of joint cartilage, and the influence of EGF on its growth and dedifferentiation. Methods: (1) Isolate chondrocytes using digestion chamber and then culture them. (2) Evaluate the digestion method by survival rate and cloning rate of isolated chondrocytes. (3) Study the influence of EGF on the growth and dedifferentiation of chondrocytes. Results: (1) Chondrocytes were totally isolated within 3 hours by the digestion chamber. (2) The survival rate was 88.4%, and the cloning rate was 29.4%. (3) EGF promoted the growth of chondrocytes ( P <0.05) . (4) Whether the EGF was added or not, dedifferentiation appeared after 8 10 times of detachment. Conclusion: The digestion chamber can make the isolation of chondrocytes more efficient. EGF can promote the growth of chondrocytes obviously, but can′t stop the dedifferentiation trend of chondrocytes.