人牙龈成纤维细胞原代培养及鉴定.docVIP

人牙龈成纤维细胞原代培养及鉴定[摘要] 目的:分离培养人牙龈成纤维细胞,为进一步研究细胞因子对牙龈成纤维细胞的调控作用提供实验条件。方法:用组织块法培养人牙龈成纤维细胞,并进行形态学和免疫学鉴定。结果:牙龈成纤维细胞原代培养成功并稳定传代,细胞呈长梭形或星形,免疫学鉴定抗波形丝蛋白抗体阳性,抗角蛋白抗体阴性,为中胚层来源的成纤维细胞。结论:组织块法培养的细胞符合牙龈成纤维细胞的形态学和免疫学特征,可作为体外细胞模型,为研究细胞因子对其功能的调控作用奠定基础。 [关键词] 牙龈成纤维细胞;细胞培养;鉴定 [中图分类号] R329.2+8[文献标识码]A [文章编号]1673-7210(2010)01(c)-026-02 The primary culture and identification of human gingival fibroblasts CUI Juan1, SUN Keqin1, LI Xia1, ZHANG Huaping2 (1.Oral Medicine Department of Oral Hospital of Shanxi Medical University, Taiyuan 030001, China; 2.Central Laboratory of Oral Hospital of Shanxi Medical University, Taiyuan 030001, China) [Abstract] Objective: To isolate and culture human gingival fibroblasts (HGFs) in vitro, and provide the experimental condition for further research on the HGFs regulation by cytokines. Methods: HGFs were primary cultured by tissue explant culture technique and identified by morphological analysis and immunocytochemical analysis. Results: The HGFs were cultured and passaged successfully. The cells showed long spindle or star appearance. By immunocytochemical analysis, these cells tested positive to anti-vimentin antibody and negative to anti-keratin antibody, indicating the cells were mesoderm derived fibroblasts. Conclusion: The cells cultured by tissue explant culture technique are consistent with typical HGFs’ characteristics of morphology and immunocytochemical observation. The HGFs are used as cell model in vitro for future research on HGFs regulation by cytokines. [Key words] Gingival fibroblasts; Cell culture; Identification 人牙龈成纤维细胞(human gingival fibroblasts,HGFs)是牙龈结缔组织中的主体细胞,在牙周组织的保护和修复中起重要作用[1]。许多学者利用牙龈成纤维细胞体外培养模型,在牙周组织发病机制及治疗方面进行了有益的探索[2]。原代培养是获取牙龈成纤维细胞的主要手段,目前有酶消化法和组织块培养法[3]。本实验采用组织块法培养人牙龈成纤维细胞,为进一步研究细胞因子对牙龈成纤维细胞的调控作用提供实验条件。 1 材料与方法 1.1 主要试剂和仪器 DMEM培养液(Gibco,美国);胎牛血清(四季青,杭州);胰蛋白酶(Sigma,美国);抗波形丝蛋白抗体、抗角蛋白抗体、免疫组化试剂盒、DAB显色试剂盒(中杉金桥,北京);25 ml塑料培养瓶、六孔板(Orange scientific,比利时);RCO3000-7 CO2培养箱(REVCO,美国);GB-Ⅱ超净工作台(北京市新技术应用研