人牙周膜成纤维细胞的原代培养及鉴定

The primary culture and identification of human periodontal ligament fibroblasts

目的体外分离培养人牙周膜成纤维细胞,进一步研究细胞因子对牙周膜细胞功能的调控作用。方法利用组织块法原代培养牙周膜成纤维细胞并传代,通过形态学及免疫细胞化学方法对其进行鉴定。结果牙周膜成纤维细胞培养成功传代,5代后细胞生长旺盛,表现为长梭形细胞,免疫细胞化学鉴定细胞抗波形丝蛋白阳性,抗角蛋白阴性,为中胚层来源的成纤维细胞。结论组织块法培养出的细胞符合牙周膜成纤维细胞的形态学及免疫学特征,生长状态良好,可作为体外的细胞模型为研究细胞因子对其功能的调控作用奠定基础。

Objective To isolate and culture human periodontal ligament fibroblasts （HPLFs） in vitro, to be used for further research on the HPLF regulation by cytokines. Methods The primary cells were isolated from human periodontal ligament and cultured by tissue explant culture technique, then passaged when the cells completely covered the bottom of culture flasks. Morphological analysis and immunocytochemical analysis were used to characterize the cell lineage. Results The HPLFs were cultured and passaged successfully. After the fifth passage, the actively proliferating cells showed long spindle appearance. By immunocytoehemieal analysis, these cells tested positive to antibodies against vimentin, and negative to antibodies against eeratin, indicating themselves to be mesoderm-derived fibroblasts. Conclusion The cells obtained by tissue explant culture technique were consistent with typical HPLFs in terms of morphology and immunocytoehemieal characteristics. The satisfactory growth condition of the cells justified their use in vitro for future research on HPLF regulation by eytokines.