骨髓间充质干细胞向脑胶质瘤定向迁移及其DAPI标记的研究

Bone marrow-derived mesenchymal stem cells to glioma directional migration and DAPI markers

目的:探讨骨髓间充质干细胞(BSMCs)向C6胶质瘤定向迁移的能力以及DAPI用于BMSCs向胶质瘤体内迁移示踪的价值。方法:直接贴壁法分离培养纯化BMSCs。利用Transwell小室建立体外迁移模型检测BMSCs向C6胶质瘤细胞定向迁移的能力。立体定向法建立大鼠C6胶质瘤模型,利用DAPI体外标记培养、纯化的BMSCs,经荷瘤侧颈内动脉灌注,观察BMSCs向C6胶质瘤组织的定向迁移能力,并评价这一过程中DAPI用于BMSCs标记的价值。结果:通过直接贴壁法分离、培养、传代,获得了纯化的BMSCs。体外迁移实验证实,BMSCs具有向C6胶质瘤细胞定向迁移的能力,迁移细胞数量为400倍视野下(32.1±10.5)/HP。DAPI标记后BMSCs细胞核呈蓝色荧光,阳性率达100%。经荷瘤大鼠颈内动脉灌注后BMSCs可以存活,并表现出向脑胶质瘤趋化迁移的特性,分布区域主要位于肿瘤内部血管周边。结论:DAPI可以用于BMSCs的体内示踪,BMSCs具有通过血肿瘤屏障向C6胶质瘤定向迁移的能力,经颈内动脉灌注是其有效的移植途径。

OBJECTIVE： To investigate the migrating potential of bone marrow-derived mesenchymal stem cells （BMSCs） towards C6 glioma and the application value of DAPI in the tracking and migration of BMSCs towards glioma in vivo. METHODS： BMSCs were isolated,cultured,passaged and purified in vitro by direct adhesion method. Using Tran- swell inserts technique,in vitro model was established and the migration of BMSCs towards C6 glioma was studied. The rat C6 glioma model was established by stereotactic procedure. After being marked with DAPI,BMSCs were injected into collateral internal carotid artery of rats bearing glioma to study their tropism for C6 glioma in vivo and evaluated the appli- cation of DAPI in this process. RESULTS： BMSCs were successive subcultured and purified by direct adhesion method. The results of in vitro migration assay showed that BMSCs could migrate through the polycarbonate filter towards C6 cells. The average number of migrating BMSCs was （32.1-I-10.5）/HP （X 400） . The nucleus of BMSCs labeled by DA- PI presented blue fluorescence,and all of the cells were labeled by DAPI. After being injected into internal carotid artery, BMSCs could survive in the brain of rats bearing glioma,and showed extensive tropism for C6 glioma and scattered around the blood vessels within the tumor. CONCLUSIONS：DAPI can be used for the tracking of BMSCs in vivo. BMSCs have the ability to penetrate blood tumor barrier and migrate towards C6 glioma,and infusion through internal carotid artery is an effective way for their transplantation.