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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 study the chimerism and distribution of allogeneic bone marrow cells in recipient organs after infusion. Methods SD rat bone marrow cells were isolated, stained with PKH26 fluorescent staining, and infused into Wistar rats through inferior vena cava or portal vein. At 0 ~ 4 days after surgery, rapamycin was injected intraperitoneally. At 7 and 14 days after transplantation of labeled bone marrow cells, blood, bone marrow, thymus, spleen, para-aortic lymph nodes and liver of the recipients were isolated and lymphocytes were isolated. The chimerism was detected by flow cytometry (FCM) Fluorescence microscopy and HE staining were used to observe the distribution of labeled cells in the lymphoid organs of the transplanted rats. Results Infusion of bone marrow cells mainly in the recipient’s lymphoid organs, differentiation and development. Over time, the number gradually decreased, eventually cleared in the spleen. FCM and fluorescence microscopy showed that the chimerism rate of labeled bone marrow cells was higher than that of the inferior vena cava through transfusion of bone marrow cells through the portal vein. Conclusion Portal vein infusion of allogenic bone marrow cells can more effectively induce the appearance of mixed chimerism in rats. In recipients, the distribution of allogeneic bone marrow cells has a corresponding rule.