人牙龈成纤维细胞原代培养及冻存和复苏

背景：牙龈成纤维细胞是牙龈固有结缔组织层中主要的细胞，在许多生理和病理过程中起着重要作用。目的：对人牙龈成纤维细胞进行原代培养、鉴定、冻存及复苏。方法：采用组织块培养法培养人牙龈成纤维细胞，并进行形态学和免疫学鉴定。对人牙龈成纤维细胞进行冻存与复苏，倒置显微镜下观察细胞形态变化。结果与结论：人牙龈成纤维细胞原代培养成功率为86.7%，细胞呈梭形或纺锤形。免疫组织化学鉴定抗波性蛋白抗体染色阳性，抗角蛋白抗体染色阴性，为中胚层来源的成纤维细胞。牙龈成纤维细胞冻存与复苏成功，细胞经2次传代后生物学性状与原代相似。提示采用组织块培养法培养原代人牙龈成纤维细胞及冻存与复苏方法可行。

Primary culture,cryopreservation and resuscitation of human gingival fibroblasts

BACKGROUND:Human gingival fibroblasts (HGFs) are the main cells in the connective tissue layers, which play an important role in many physiological and pathological processes.OBJECTIVE:To perform primary culture, identification, cryopreservation and resuscitation for HGFs.METHODS:First, the HGFs were cultured by the methods of tissue culture and identified by morphological and immunocytochemical analysis. Next, HGFs were frozen and resuscitated. And then their morphological changes were observed by the inverted microscope.RESULTS AND CONCLUSION:The success rate of HGFs from the primary cells was 86.7%, and the HGFs showed fusiform or spindle-shaped. The immunochemistry study indicated that vimentin showed positive staining; while cytokeratin showed negative staining, which originated from mesoblastma. The cryopreservation and resuscitation of HGFs were successful. The biological character of HGFs after second passage was similar to that of the original generation. These results suggest that the primary culture, cryopreservation and resuscitation of HGFs can be feasible by using the method of tissue culture.