用猪AMI模型研究荧光染料CFSE和Brdu作为移植细胞标记物的实用性

目的研究荧光染料CFSE和Brdu作为体内细胞移植标记物的实用性.方法构建猪急性心肌梗死(AMI)模型.用CFSE和Brdu对骨髓单个核细胞进行双标记,经冠状动脉回输至MI区.4周后,利用激光共聚焦显微镜和免疫组织化学法分别观察CFSE和Brdu的定位和标记情况.结果细胞移植4周后,发现Brdu阳性细胞多位于心肌疤痕形成区且MI区的新生血管内壁也出现阳性染色.在MI区可见CFSE标记的带绿色荧光的细胞,但背景色较高.结论在动物实验中,CFSE和Br-du均可作为移植细胞标记物,Brdu的标记效果优于CFSE,适于同时进行形态学和组织学研究.

The study on utilities of CFSE and Brdu as cellular labels in vivo in porcine myocardial infarction model

Objective To evaluate the utilities of bromodeoxyurine (Brdu) and vital dye 5,6-carboxyfluorescein diacetate succinimidyl ester(CFSE) as cellular labels in vivo.Methods A myocardial infarction was created and PBMCs were separated and labeled with Brdu and CFSE.We injected the labeled cells to the infraction region through coronary artery.After four weeks of cell transplantation,the precise location of CFSE-labeled pBMCs was observed under laser scanning confocal microscope (LSCM) and by immunohistochemical staining. The positive signals of the Brdu-labeled cells was accessed.Results After four weeks of implantation by LSCM,fluorescence signal was detected,but a high background occurred and immunohistochemical staining for Brdu in myocardial scar tissue was also detected Some endothelial cells in blood vessels of transplanted region were stained positively for Brdu.Conclusion CFSE and Brdu can be used in several aspects in medical research,especially as in vivo indicators of cell graft.The positive signals from Brdu is optimum in the morphological and histological study.By contrast,fluorescence signals from CFSE is also intense,and yet the positive results will be interfered by high background.