四川大学学报（医学版）Issue(2)：266-270,5.
大鼠骨髓间质干细胞的BrdU体外标记和体内示踪研究
In vitro Analysis and in vivo Tracing of BrdU-labeled Rat Bone Marrow Mesenchymal Stem Cells
摘要
Abstract
Objectives To investigate the optimal condition of bromodeoxyuridine (BrdU) labeling for bone marrow mesenchymal stem cells (BMSCs) in vitro and the feasibility of in vivo tracing of BrdU-labeling BMSCs. Methods BMSCs were isolated from Wistar rats and were in vitro routinely cultured. The third passage BMSCs was used for identification of special surface antigens by immunohistochemical methods. The purified BMSCs were incubated with BrdU at different concentrations for different incubating time to investigate optimal BrdU concentration and incubating time for cell labeling. The cell labeling index of BrdU was calculated with immunohistochemical analysis. BMSCs labeled BrdU were injected into damaged gastric mucosa of rats by micro injector. The colonization of BMSCs labeled BrdU in gastric mucosa was viewed. Results After purification and proliferation, the primary cultured BMSCs were uniformly long spindle-shapped form and formed cell colony, which showed the characteristics of stem cell. Immunocytochemistry showed BMSCs were positive for CD44 and CD90, while negative for CD14, CD45. The labeling rate of BrdU increased with the labeling time lasting and reached its height at 48 h. After incubating 48 and 72 hours, the labeling rate of BrdU with a concentration of 10 μmol/L was higher than that of BrdU with a concentration of 5 μmol/L (P < 0. 05) and similar with that of BrdU with a concentration of 15 μmol/L (P>0. 05). In addition, the BrdU labeling could be detected after five consecutive passages and the labeling time could keep 21 d. The pathological observation demonstrated that BrdU-labeled BMSCs could colonize the damaged gastric mucosa with normal morphologic characteristics during observation period. Conclusion BrdU labeling might be a feasible method for dynamic observation of the migration, growth and differentiation of migrating BMSCs in colonizing sites.关键词
5-溴脱氧尿嘧啶核苷/骨髓间质干细胞/细胞培养Key words
Bromodeoxyuridine/Bone marrow mesenchymal stem cells/Cell culture引用本文复制引用
张明鸣,贾贵清,程惊秋,杨平,陆燕蓉,伍晓汀..大鼠骨髓间质干细胞的BrdU体外标记和体内示踪研究[J].四川大学学报（医学版）,2012,(2):266-270,5.基金项目
国家自然科学基金(批准号30672065)、教育部博士点基金(批准号20070610146和20090181120107)和四川大学青年科学基金(2008096)资助 （批准号30672065）
