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目的采用羧基荧光素二醋酸盐琥珀酰亚胺酯(carboxyfluorescin diacetate succinimidyl ester,CFSE)与溴化脱氧尿嘧啶核苷(5-bromo-2-deoxyuridine,Brdu)两种细胞标记物观测肝移植术后受体骨髓间充质干细胞(MSCs)肝内分布情况,并对两种方法进行比较。方法在已构建的DA-Lewis大鼠原位肝移植模型及提取培养Lewis大鼠MSCs的基础上分别用CFSE和Brdu对Lewis来源的MSCs进行标记,术中经门静脉输注,通过荧光显微镜和免疫组织化学法观测术后2mo内各时间点受体肝组织内MSCs的分布情况。结果CFSE细胞标记率约为(94.1±1.4)%。受体肝组织第1、7、14天内有CFSE标记的MSCs聚集,对照组第5天肝组织内发布的CFSE标记MSCs消失。BrdU细胞标记率约为(90.3±3.5)%。受体肝组织第14天、1个月、2个月可见Brdu阳性的MSCs分布,对照组相应时间点肝组织内未发现Brdu阳性的MSCs。结论CFSE和Brdu均为MSCs的良好标记物。CFSE标记细胞后荧光迅速减弱,适用于短期示踪。Brdu标记细胞后可维持较长时间,适用于长期示踪。两种标记方法均是MSCs较为简单有效的方法,实验显示受体MSCs大部分分布于移植肝脏。
OBJECTIVE: To observe the effects of liver transplantation by using carboxyfluorescin diacetate succinimidyl ester (CFSE) and 5-bromo-2-deoxyuridine (Brdu) Post-receptor bone marrow mesenchymal stem cells (MSCs) in the liver distribution, and compare the two methods. Methods Based on the established model of orthotopic liver transplantation in DA-Lewis rats and the culture of Lewis rat MSCs, Lewis derived MSCs were labeled with CFSE and Brdu, respectively. Intravenous transfusions were performed by fluorescence microscopy and immunofluorescence Histochemistry was used to observe the distribution of MSCs in the recipient liver tissue at each time point within 2 months. Results The CFSE cell labeling rate was (94.1 ± 1.4)%. CFSE-labeled MSCs were collected on the 1st, 7th and 14th day in the recipient liver tissues, while the CFSE-labeled MSCs released on the 5th day in the control group disappeared. BrdU cell labeling rate was about (90.3 ± 3.5)%. Brdu-positive MSCs were observed on the 14th, the 1th and the 2th month in the recipient liver tissues, and no Brdu-positive MSCs were found in the liver of the control group at the corresponding time points. Conclusion Both CFSE and Brdu are good markers of MSCs. CFSE labeled cells rapidly weakened fluorescence, suitable for short-term tracing. Brdu labeled cells can be maintained for a long time, suitable for long-term tracer. Both labeling methods are relatively simple and effective methods of MSCs. Experiments show that most of the recipient MSCs are distributed in the liver graft.