异基因骨髓细胞在大鼠体内嵌合分布特性的研究  

Distribution features of chimerism induction of allogeneic bone marrow cells in rats

作　　者：孙佩[1,2] 高庆贞[2] 任万军[2] 张瑞斌[2] 王小平[2] 刘子栋[2] 

机构地区：[1]山东大学医学院,济南250012 [2]山东大学附属济南市中心医院,济南250013

出　　处：《山东大学学报（医学版）》2010年第1期38-40,44,共4页Journal of Shandong University:Health Sciences

基　　金：济南市科技局科技明星计划项目(98122)

摘　　要：目的研究大鼠异基因骨髓细胞输注后在受者器官的嵌合状态及分布规律。方法分离SD大鼠骨髓细胞,用PKH26荧光染色标记,通过下腔静脉或门静脉输注到Wistar大鼠。在手术的0～4d,腹腔注射雷帕霉素。在输注标记骨髓细胞术后7、14d,取受者大鼠的血液、骨髓、胸腺、脾脏、腹主动脉旁淋巴结、肝脏,分离淋巴细胞,用流式细胞仪(FCM)检测嵌合情况;荧光显微镜和HE染色观察标记细胞在移植大鼠淋巴器官内的分布情况。结果输注的骨髓细胞主要分布在受者的淋巴器官内,进行分化和发育。随时间的延长,数量逐渐减少,最终在脾脏被清除。FCM和荧光显微镜检测显示,通过门静脉途径输注骨髓细胞,标记骨髓细胞的嵌合率高于下腔静脉途径。结论门静脉途径输注异基因骨髓细胞能更有效地诱导大鼠混合性嵌合状态的出现;在受者体内,异基因骨髓细胞的分布有相应的规律。Objective To investigate the distribution and the chimerism features after infusion of allogeneic bone marrow cells （BMCs） in rats. Methods BMCs of SD rats were PKH26 labeled and 5×10^7 cells were infused to Wistar rats via the portal or inferior vena cava. Rapamycin was injected daily from day 0 to 4 after surgery. On day 7 and 14,the recipient rats were sacrificed and the blood,the BMCs,the thymus,the livers,the spleens,and the lymph nodes were collected. The lymphocytes of these organs were separated and then were analyzed by flow cytometry to detect the PKH26 labeled BMCs. The smear of peripheral blood mononuclear cells,the smear BMCs,and frozen tissue sections of the thymus,lymph nodes,the livers,and the spleens were observed with a fluorescence microscope. Results Both flow cytometry and fluorescence microscopy demonstrated that BMCs injection induced mixed chimerism in adopted Wistar rat organs of the thymus,the liver,the spleen,and the lymph node. The chimerism level was higher when injected via the portal vein than via the inferior vena cava. Fluorescence microscopy detected the distribution features of PKH26 labeled cells in those organs,and indicated that donor BMCs can differentiate in those organs and be finally eliminated in the spleens. Conclusion BMCs infusion can induce mixed chimerism in recipient organs with different distribution features,and portal vein infusion is a more effective strategy than inferior vena cava infusion.

关 键 词：骨髓细胞 门静脉 免疫耐受 嵌合状态 大鼠 

分 类 号：R551.3[医药卫生—血液循环系统疾病] R-332[医药卫生—内科学]