目的研究猪耳软骨细胞在体外培养传代过程中,细胞老化相关基因和胞外基质相关基因表达水平的改变。方法取体外培养猪耳软骨细胞,通过光镜,对各代细胞的形态学变化进行观测。RT-PCR检测各代细胞中bcl-2、端粒酶、Ⅰ型胶原和Ⅱ型胶原在mRNA表达水平上的改变。结果体外培养软骨细胞随着老化的进程,逐渐显现衰老和凋亡的特征。凋亡抑制基因bcl-2和控制细胞分裂的端粒酶的表达显著下降(P<0.01)。软骨细胞的特征性基质分子Ⅱ型胶原在第3代时表达水平有显著下降(P<0.01),降至原代细胞的5.47%±1.04%,在第4代几乎不再表达。而Ⅰ型胶原在第5代时,有显著上升(P<0.01);在第9代时又显著下降(P<0.01)。结论猪软骨细胞经体外培养,由第4代起逐渐发生去分化,丧失软骨细胞的表型。同时,细胞的增殖分裂能力亦逐步减退,细胞凋亡现象明显增加。 OBJECTIVE To study the changes of cell-related genes and extracellular matrix-related genes during the passage of porcine ear ear cartilage cells in vitro. Methods Porcine ear cartilage cells were cultured in vitro, and morphological changes of each passage were observed by light microscope. The changes of mRNA expression of bcl-2, telomerase, type I collagen and type II collagen were detected by RT-PCR. Results Chondrocytes cultured in vitro gradually showed the characteristics of aging and apoptosis with the progress of aging. The expression of apoptosis-suppressing gene bcl-2 and cell division controlling telomerase were significantly decreased (P <0.01). The expression of collagen Ⅱ, a characteristic matrix molecule of chondrocytes, decreased significantly at the third passage (P <0.01), decreased to 5.47% ± 1.04% of the primary cells, and almost no longer expressed at the fourth passage. Collagen type Ⅰ increased significantly (P <0.01) at the fifth generation and decreased significantly at the ninth generation (P <0.01). Conclusion Pig chondrocytes were cultured in vitro and gradually dedifferentiated from the fourth passage to lose the phenotype of chondrocytes. At the same time, cell proliferation and division ability also gradually decreased, the phenomenon of apoptosis increased significantly.