骨髓间充质干细胞在致敏小鼠体内的示踪归巢实验

Bone Marrow Mesenchymal Stem Cells Homing in Sensitized Mice

【目的】了解骨髓间充质干细胞在以异基因脾细胞输注致敏小鼠体内的归巢情况。【方法】正常同种异基因小鼠骨髓间充质干细胞的体外培养及鉴定后,体外予羧基荧光素二醋酸盐琥珀酰亚胺酯荧光染料标志,经尾静脉注射到致敏组和非致敏组小鼠体内(每组10只),在6 h、24 h和48 h处死动物,分别应用冰冻切片和流式检测方法了解骨髓间充质干细胞在各个脏器的分布情况。【结果】冰冻切片和流式检测结果均表明骨髓间充质干细胞在致敏小鼠体内迅速和持久在脾、肝脏聚集,而非致敏动物却没有明显聚集到肝脾等脏器,而是最终归巢到了骨髓。其中流式细胞仪定量结果表明:6 h时致敏小鼠脾脏荧光阳性细胞为(3.78±0.66)%,与同期正常小鼠脾脏结果比较P<0.05,有统计学意义;而在48 h时致敏小鼠骨髓荧光阳性细胞为(0.43±0.35)%,同期正常小鼠骨髓为(5.40±1.58)%,两者比较P<0.05,有统计学意义。【结论】应用异基因脾细胞致敏的小鼠体内肝脾等免疫脏器明显受到致敏影响,其对骨髓间充质干细胞体内归巢的定位具有很高的趋化吸附作用,中止了骨髓间充质干细胞正常状态下向骨髓的归巢。

[Objective] To explore the homing characteristics of bone marrow mesenchymal stem cells （MSCs） in the allogeneic splenocytes sensitized mice. [ Method ] After normal allogene bone marrow MSCs were cultured and evaluated, MSCs were labeled by 5-and 6-carboxyfluorescein diacetate succinimidyl ester （CFSE） in vitro. After injected CFSE labeled MSCs （CFSE ＋ cells） through vena caudalis in 6,24, and 48 h,homing trace were explored in the different organs both in the sensitized and normal mice group （each group 10 samples mice） through frozen section and flow cytometry （FCM） detection methods. [Results] Both in the methods of frozen section and FCM, we discovered in the sensitized mice group MSCs were aggregated and homing in spleen and liver quickly and lastingly, while in the normal mice group MSCs were homing to the bone marrow in the end. Among those FCM quantitative result, in 6 h the percentage of spleen CFSE ＋ ceils in the sensitized mice group was （3.78＋0.66）%, significant difference was found in spleen between the sensitized and normal group at the same time （P 〈0.05）; while in 48 h, the percentage of bone marrow CFSE＋ cells in the sensitized and normal mice group were （0.43-＋0.35）% and （5.40-＋1.58）%, respectively （P〈 0.05）. [Conclusion] Immunity organs such as liver and spleen, influenced by the sensitized of allogeneic splenocyted transfusion, had the orientating function in the normal allogene bone marrow MSCs homing, which interrupted the homing to the bone marrow in the normal condition.