摘要
目的探讨组织块法培养大鼠颌下腺细胞(RSGC)的方法与细胞生长的特点。方法采用组织块法培养RSGC,使用酶消化法和差速贴壁法进行细胞纯化,免疫组织化学SABC方法进行细胞角蛋白-8免疫组化染色,进行细胞来源鉴定。倒置显微镜下进行形态学观察,取第2代RSGC,FDA-PI双色荧光法检测细胞活力,MTT法绘制生长曲线。结果细胞角蛋白-8染色阳性。倒置显微镜下,细胞主要呈3种形态,为圆形亮细胞、多角形暗细胞、梭形暗细胞。细胞活力>95%,生长曲线表示细胞5d开始进入对数生长期。结论此方法简便,易操作,获得了大量的RSGC,为颌下腺细胞作为种子细胞的颌下腺组织工程研究提供了实验基础和理论依据。
Objective To explore the application of tissue-explant technique in culturing rat submandibular gland cell （RSGC）and the characteristics of RSGC. Methods RSGCs were cultured using tissue-explant technique. The cells were purified by enzymatic digestion and differential adhesion. The cell phenotype was identified by cytokeratin-8 immunohistochemical staining. Cellular morphology was observed and photographed under inverted microscope. The cell viability and growth were determined by a double-staining procedure using FDA-PI and MTT assay,respectively. Results Cytokeratin-8 was positive stained in the immunohistochemical staining. The cell viability was more than 95%. The cell growth curve showed that RSGCs were in logarithmic phase since day 5. Conclusion Tissue-explant technique is an easy way to purify plentiful RSGC with normal functions,and it can be used in further research of tissue engineering of submandibular gland.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2010年第3期194-196,共3页
Journal of China Medical University
基金
辽宁省教育厅高等学校科研项目(2008843)
关键词
颌下腺细胞
组织块培养方法
酶消化法
差速贴壁法
组织工程
submandibular gland cell
tissue-explant technique
enzymatic digestion
differential adhesion
tissue engineering
