人牙周膜成纤维细胞分离、纯化培养及生物学特性观察

The culture of periodontal ligament fibroblasts and study on their biological behaviors in vitro

采用胶原酶及胰蛋白酶联合消化法 ,简便、快速地获得大量成活率高的人牙周膜细胞。在 DMEM培养基中进行了原代及传代培养 ,对其中的成纤维细胞进行分离纯化。相差显微镜下 ,牙周膜成纤维细胞呈星形或长梭形 ,免疫组化波丝蛋白阳性 ,角蛋白阴性 ,证实该细胞来源可靠。酶消化法不失为一种简便、快速、可靠的细胞原代分离培养方法。

To investigate the behavior of PDL Fibroblasts in vitro, the periodontal ligament fibroblasts were harvested from orthodontic extracted premolars subjected with sequential digestion with 0.25% trypsin and 0.2% collagenase. The isolated cells were cultured in DMEM medium and PDL fibroblasts were purified in subculture. The PDL fibroblasts were identified by positve stain for vimentin and negative for keratin. Cell morphology and growth pattern were observed. The results revealed that PDLF cells in this culture system kept their phenotype in vitro.