同种异体血清体外培养兔膝关节软骨细胞的增殖

摘要

背景:兔关节软骨细胞体外单层培养采用胎牛血清培养基易去分化,有必要寻找合适培养基来提高兔关节软骨细胞培养质量.目的:观察同种异体兔血清对体外培养的兔膝关节软骨细胞增殖能力的影响.方法:以0.4%链霉蛋白酶和0.025%Ⅱ型胶原酶分离成年兔膝关节软骨细胞,将获得的软骨细胞随机分为实验组和对照组.实验组以体积分数10%异体兔血清+DMEM/F12 培养;对照组以体积分数10%胎牛血清+DMEM/F12 培养,传代培养至4代.结果与结论:实验组前4 代软骨细胞增殖较对照组慢,但软骨细胞形态未发生明显改变而对照组软骨细胞出现去分化现象.提示体积分数10%异体兔血清培养利于维持软骨细胞增殖和形态的稳定,是较好的获取大量优良软骨细胞的体外培养方式.%BACKGROUND: Because chondrocytes are easy to dedifferentiate when they are cultured in vitro with fetal bovine serum, it is necessary to find a suitable medium to improve the quality of rabbit articular chondrocytes. OBJECTIVE: To observe the effects of allogeneic rabbit serum on the proliferation of rabbit articular chondrocytes cultured in vitro. METHODS: The chondrocytes were isolated from rabbit knee articular cartilage tissues with 0.4% Pronase and 0.025% collagenase , then equal randomly divided into two groups: the chondrocytes of the experiment group were cultured in ⅡDulbecco's modified Eagle's medium/Ham's nutrient mixture F-12 containing 10% allogeneic rabbit serum, whereas the culture medium of 10% fetal bovine serum and Dulbecco's modified Eagle's medium/Ham's nutrient mixture F-12 was used in control group. After subculture, passage 4 cells were obtained. RESULTS AND CONCLUSIONS: Compared with the control group, the chondrocytes in the experiment group proliferated slowly before the 4th passage. But at the same time, the morphology of chondrocytes in the experiment group was better than that in the control group. When the chondrocytes were at the fourth passage, we found that the shape and growth of chondrocytes in the experiment group achieved better results. Meanwhile the dedifferentiated chondrocytes could be observed in the control group. The 10% allogeneic rabbit serum medium is good for maintaining the shape and proliferation of chondrocytes, so it is indicated that we can get a large number of good chondrocytes through this way.