下颌骨髁突软骨细胞传代过程中生物学特性变化

Phenotypic Changes of Mandibular Condylar\;Cartilage Cells During Subculture

目的:观察体外培养及传代对人髁突软骨细胞生物学特性的影响。方法:采用体外细胞培养、免疫生化及分子生物学方法观察传代过程中人髁突软骨细胞形态、型胶原及蛋白多糖合成及、、型前胶原的mRNA水平变化。结果:传代过程中人髁突软骨细胞型胶原合成及其mRNA水平逐渐降低,至第7代已基本丧失。而、型胶原的mRNA水平随传代逐渐升高,同时伴有细胞蛋白多糖合成的减少,细胞壁形态由圆形或多角形转变为长梭形占优势。结论:体外培养及反复传代可影响软骨细胞的表型特征。髁突软骨细胞的反分化标准应从胶原。

Objective:To demonstrate the phenotypic changes of mandibular condylar cartilage(MCC) cells during subculture in vitro. Methods: MCC cells were harvested from human fetus by digestion of collagenase and cultured in DMEM supplemented with 10% newborn calf serum, then they were passaged when reached confluence.Morphological changes were observed under phase contrast microscope. Type Ⅱ collagen, proteoglycan synthesis and type Ⅰ,Ⅱ and Ⅲ procollagen mRNA levels were also studied in serial monolayer cultures. Results: During subculture,type Ⅱ collagen and proteoglycan synthesis were decreased, being accompanied by a lower type Ⅱ collagen mRNA level. In contrast, type Ⅰ and Ⅲ procollagen mRNA levels were elevated progressively. The morphology of polygonal shaped chondrocytes was lost to the bipolar fibroblastic cells. Conclusion: Passage and culture can affect the phenotypic property of MCC cells.\;