摘要
目的:分离获取大鼠耻骨联合软骨细胞,体外培养并进行形态学观察。方法;采用机械及胰蛋白酶、胶原酶序贯消化法,从大鼠耻骨联合软骨组织中分离出软骨细胞,用含15%FBS的DMEM/F12培养液原代和传代培养,倒置显微镜下动态观察细胞形态及生长情况,阿尔辛蓝组织化学、Ⅱ型胶原免疫组化染色鉴定,通过扫描及透射电镜观察培养细胞的超微结构。结果:体外培养的大鼠耻骨联合软骨细胞呈短梭形、三角形,有短细胞突起,细胞浆内质网发达,线粒体丰富,富含分泌颗粒。细胞胞浆糖胺聚糖、Ⅱ型胶原染色均阳性。结论:耻骨联合细胞体外培养过程中,较好地保持了生长期软骨细胞的形态特征。
Objective:To explore the morphological charaters of rat pubic symphysis cartilage cells (PSCC) cultured in vitro. Method: The chondrocytes were obtained from rat pubic symphysis cartilage by mechanical and trypsin-collagenase digested. Phase contrast microscope, alcian blue dyeing, collagen type Ⅱ immunocytochemical stain, scanning electron microscope (SEM) and transmission electron microscope (TEM) were used to observe the morphological characters of PSCC. Result: The phase contrast micrography and SEM showed that PSCC cultured in vitro underwent fusiform, spindle- shaped, polygonal shape. TEM indicated that PSCC had chondrocyte-like phenotype: with prosperous endoplasmic reticulum, mitochondrion and secretory granule. There were glycosaminoglycan and collagen type Ⅱ in the plasm showed by cytochemistry stain. Conclusion: PSCC cultured in vitro retain the main morphological character of chondrocyte in the period of growth.
出处
《临床口腔医学杂志》
2005年第11期662-664,共3页
Journal of Clinical Stomatology
基金
广东省重大科技专项基金资助项目(2004A30201004)
关键词
耻骨联合
软骨细胞
细胞培养
形态学
pubic symphysis
chondrocyte
cell culture in vitro
morphology