骨髓基质细胞经CM-Dil标记后的活性及移植到损伤脊髓的示踪观察

目的 观察骨髓基质细胞(BMSCs)经CM-Dil荧光染料标记后的生物活性及移植后在损伤脊髓中的迁移和分布.方法 取Wistar大鼠的第3代BMSCs消化后分两组,实验组和对照组,实验组以CM.DiI标记,另一组正常细胞不标记,测定生长曲线.以改良Allen法制备脊髓损伤模型,将CM.DiI标记的BMSCs通过蛛网膜下腔注射移植,分别于移植后24 h、1、2、3、4周取损伤脊髓段作冰冻切片,倒置荧光显微镜下观测移植的BMSCs在损伤脊髓中的迁移和分布规律.结果 BMSCs经CM-Dil荧光染料标记成功率100%,实验组和对照组的生长曲线基本吻合;移植后24 h,脊髓损伤区未见标记细胞,移植后1周,标记的BMSCs开始迁移到损伤区硬脊膜下,移植后2～3周.BMSCs迁移、聚集在损伤脊髓的中心,移植后4周,仍有标记细胞聚集在脊髓损伤中心.结论 (1)BMSCs经CM-Dil荧光染料标记后的生物活性保持正常;(2)CM-Dil标记的BMSCs移植后能迁移聚集在损伤脊髓的中心,CM-Dil在活体内至少能维持4周

Activity and migration of bone marrow stromal cells labeled by CM-Dil in injured spinal cord

Objective To study the activity of CM-Dil labeled bone marrow stromal cells and observe the migration and distribution of BMSCs in injured spinal cord with intradural space transplantation. Methods The growth curve of CM-Dil labeled BMSCs was assessed and compared the with that of the unmarked BMSCs. The spinal cord injury model was prepared according to the modified Allen method. The labeled BMSCs were transplanted into intradural space apart from the injured spinal cord. Spinal cord was dissected at 24 h,1,2,3 and 4 weeks after cells transplantation. Four μm thick sections were cut on a cryostat and observed under the fluorescence microscopy. Results The activity of CM-Dil labeled BMSCs was normal. One week after transplantation, BMSCs were mainly located under the dura mater. Two to 4 weeks later,BMSCs migrated and concentrated into the injured spinal parenchyma. Conclusion CM-Dil labeled BMSCs could migrate to the injured parenchyma of spinal cord and kept red fluorescent for 4 weeks.