两种体外培养人牙周韧带成纤维细胞方法比较  

Comparison of two culture methods of human periodontal ligament fibroblasts in vitro

作　　者：王晓丽[1] 宋萌[1] 钮晓勇[1] 娄佳宁[1] 

机构地区：[1]上海交通大学附属第一人民医院口腔科,上海200080

出　　处：《口腔医学》2008年第3期157-160,共4页Stomatology

摘　　要：目的探索一种在体外短时间内简便、可靠获取大量人牙周韧带成纤维细胞（human periodontal ligament fibroblast，HPLF）、建立稳定的体外培养体系的方法。方法采用酶消化法和组织贴块法进行HPLF体外原代培养及传代培养的对比研究。通过细胞形态学、超微结构观察及波形蛋白和角蛋白免疫组化染色等对细胞进行定性研究；测定细胞生长曲线了解细胞生长基本规律及其增殖能力。结果采用酶消化法和组织贴块法均可成功的进行HPLF连续传代培养。最高传代数为30代。培养的细胞具有成纤维细胞的典型形态，波形蛋白染色阳性，角蛋白染色阴性，生长稳定期倍增时间为48～72h。组织块培养法需培养时间长，原代培养获取的细胞量较少，较难传代。酶消化法可短时间内获取大量细胞，细胞产量高，但操作手续复杂易污染，细胞易受损伤。结论成功建立了一个稳定的HPLF体外培养体系。除常用的组织块法外，胰蛋白酶及胶原酶联合消化法不失为一种简便、快速、可靠的组织原代分离培养方法。Objective To compare the two methods of culturing normal human periodontal hgament fibroblasts （HPLFs） in vitro. Metllmds Primary and secondary HPLFs were cultured in vitro using the enzyme digestion method and the tissue block method. The HPLFs were identiffed by morphological study, transmission electron microscope and immunohistochemistry method. The cell growth curve was determined. Results Both the culture methods could do successfid passage culture. The highest passage number was 30. The cultured HPLFs had the typical property of the fibroblast cell. It showed the positive staining of the vimentin（ ＋ ） and the negative staining of cytokemtin（ - ）. The doubling time of growth was 48 - 72h. The HPLFs culture using the enzyme digestion method was more simple, stable and faster than the tissue block method. Conehtsion The HPLFs culture using the enzyme digestion method is a simple, rapid and stable method for culturing HPLFs in vitro

关 键 词：牙周韧带 牙周韧带成纤维细胞 细胞培养 

分 类 号：R780.2[医药卫生—口腔医学]