中文摘要：

目的探讨最佳的生物标记方法示踪细胞，更好地观察细胞移植后在体内的归巢情况。方法本实验室用一种新型慢病毒（eGFP＋PUROLentivirus）转染了四种细胞，确定了四种细胞的最佳感染复数，细胞转染成功后，用最佳浓度的嘌呤霉素筛选，获得四种稳定的GFP阳性表达细胞株。所用的细胞包括人脐带间充质干细胞（hUC—MSC），树齁脐带间充质干细胞（TS—UC-MSC），人胚肾细胞（293T），C57BL小鼠骨髓间充质干细胞（C57．BMSC）。结果用新型慢病毒（eGFP＋PUROLentivirus）成功转染四种常用细胞，转染成功率100％，细胞成功标记上GFP荧光，并确定了最佳的嘌呤霉素使用浓度，确定了不同细胞的感染复数。最终确定TS—UC-MSC的最佳感染复数为240，hUC—MSC的最佳感染复数为150，293T的最佳感染复数为100，C57-BMSC的最佳感染复数为50。用带荧光的C57一BMSC细胞回输C57BL小鼠体内，冰冻切片在肝中找到标记细胞，在脾、肺、肾、心未找到标记细胞，说明外来细胞主要在肝脏中代谢。结论在细胞移植治疗时，细胞成功标记上GFP能在动物体内进行示踪，良好的细胞示踪方法可以很好地反映所研究细胞在体内外的动向。

英文摘要：

Objective Explore the best biomarker method to track cells and better observe the homogenization of cells after transplantation in the body. Method Four cells were transfected with a novel lentivirus （eGFP＋PURO Lentivirus）. The optimal MOI of the four cells was determined. After the cells were successfully transfected, the cells were screened with the optimum concentration of puromycin. Four stable GFP-positive cell lines were obtained. The cells used included human umbilical cord mesenchymal stem cells （hUC-MSC）, tree shrew umbilical cord mesenchymal stem cells （TS-UC-MSC）, human embryonic kidney cells （293T）, C57BL mouse bone marrow mesenchymal stem cells （C57-BMSC）. Results Four kinds of common cells were successfully transfected with novel lentivirus （eGFP ＋ PURO Lentivirus）. The success rate of transfection was 100 %. The cell was successfully labeled with GFP fluorescence and the optimal concentration of puromycin and MOI value was determined. The optimal MOI of TS-UC-MSC was 240, the optimal MOI of hUC-MSC was 150, and the optimal MOI of 293T was 100, the optimal MOI of C57-BMSC was 50. C57BL mice were transfused with C57-BMSC cells labeled by GFP. Frozen sections showed labeled cells were in the liver. The labeled cells were not found in the spleen, lung, kidney and heart, indicating that the foreign cells were mainly metabolized in the liver. Conclusion In cell transplantation treatment, the cells successfully labeled GFP can be traced in animals, a good cell tracer method can be a good reflection of the study of cells in vitro and in vivo trends.