Ⅱ型胶原酶消化时间对兔软骨细胞生物学特性的影响

The impact of collagenaseⅡdigestion time on the biological characteristics of rabbit chondrocytes

目的探索Ⅱ型胶原酶消化法分离兔肋软骨细胞的最佳消化时间。方法取3月龄新西兰大白兔肋软骨组织,采用0.2%Ⅱ型胶原酶分别消化4、8、12、16、20h,对应为A、B、C、D、E组,获取细胞后进行培养。通过比较5组P2代软骨细胞在形态、增殖速度、细胞内基质分泌等方面的差异,选择合适的消化时间。结果 C、D、E组获取的原代软骨细胞数量比A、B组多;C、D组P2代软骨细胞增殖能力强于其他3组;C、D组P2代细胞内基质分泌情况最佳。结论Ⅱ型胶原酶最佳消化时间为12-16h,获取的原代软骨细胞数量多,活性率较高,增殖能力强,P2代软骨细胞表型良好。

Objective To investigate the best digestion time for rabbit rib cartilage cells separation by collagenaseⅡ digestion method.Methods Rabbit cartilage tissue was taken from 3-month-old New Zealand white rabbits,and then digested in 0.2%collagenaseⅡ solution for 4,8,12,16 and 20hunder vibration at 37 ℃,respectively.The corresponding experiments were named as group A,B,C,D,and E.The obtained primary cells were cultured and passaged after counting.The cell morphology,proliferation rate,secretion of extracellular matrix of passage 2cells were compared to chose the best digestion time.Results The primary chondrocytes obtained in group C,D and E were more than the other two group A and B.The chondrocytes cytoactive in group C and D were better than the other three group A,B and E.The passage 2chondrocytes in group C,D had better secretion of extracellular matrix capability than the other three groups.Conclusion The best collagenaseⅡ digestion time is 12 to 16h,at this digestion time period,more primary chondrocytes with high cytoactive and strong proliferation capability can be obtained,and the phenotype of their passage 2cells is quite good.