人牙龈成纤维细胞原代培养及冻存和复苏  

Primary culture,cryopreservation and resuscitation of human gingival fibroblasts

作　　者：惠敏[1] 倪莹[2] 郑静[3] 刘健[4] 王帅[5] 张晓明 

机构地区：[1]滨州医学院附属医院口腔内科,山东省滨州市256600 [2]滨州医学院附属医院放射科,山东省滨州市256600 [3]滨州医学院附属医院口腔修复与正畸科,山东省滨州市256600 [4]滨州医学院口腔学院,山东省滨州市256603 [5]滨州医学院临床试验中心,山东省滨州市256603 [6]日照市中医院口腔正畸科,山东省日照市276800

出　　处：《中国组织工程研究》2012年第46期8620-8624,共5页Chinese Journal of Tissue Engineering Research

基　　金：滨州医学院科技计划研究课题(BY2011KJ073)~~

摘　　要：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。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.

关 键 词：人牙龈成纤维细胞 细胞培养 冻存 复苏 鉴定 组织构建 

分 类 号：R318[医药卫生—生物医学工程]