CM-DiI示踪人脐血源基质细胞体内迁移定位的研究

Immigration and localization of human umbilical cord blood-derived stromal cells:Traced by CM-DiI in vivo

目的:课题组前期实验发现人脐血源基质细胞(hUCBDSC)体外具有促进造血细胞集落扩增的能力,文中拟采用CM-DiI荧光标记技术,观察hUCBDSC移植后的归巢、定位和增殖情况。方法:传代培养hUCBDSC,CM-DiI荧光染料预染后经尾静脉输入BALB/c-nu/nu裸鼠体内,分别于移植后1、7、14、21 d取裸鼠骨髓、脾脏、肝脏、肺脏组织,激光共聚焦显微镜观察CM-DiI标记hUCBDSC体内分布情况。结果:传代培养hUCBDSC呈成纤维样,CM-DiI染色后胞膜呈红色。经尾静脉移植至裸鼠体内,移植后1 d,hUCBDSC广泛分布在骨髓、脾脏、肝脏、肺脏等组织中,移植7 d以后,hUCBDSC主要分布在骨髓,在骨髓中增殖、分化;脾脏、肝脏、肺脏等组织中的hUCBDSC明显减少。结论:人脐血源基质细胞经尾静脉输注可"归巢"至骨髓,并在骨髓中增殖、分化,重建受损造血微环境。

Objective： Our group has studied human umbilical cord blood-derived stromal cells （hBMSC） for many years and found that hUCBDSCs could promote the amplification of hematopoietic cell colony as compared with human bone marrow stromal cells （hBMSCs）. This study aims to trace the immigration, localization and proliferation of hUCBDSCs in vivo. Methods ： Subcuhured hUCBDSCs were stained by DMDiI and then infused into post-radiated BALB/c-nu/nu nude mice. The CM-DiI dyed hUCBDSCs in the bone marrow, spleen, liver and lungs were observed by confocal microscopy at 1, 7, 14 and 21 d after transplantation. Results： The hUCBDSCs after serial subculturing appeared fibroblast-like, the cell membrane red after dyed by CM-DiI. After a transient retention in the liver, spleen and lungs, the hUCSDSCs moved quickly to the bone marrow, and proliferated and differentiated there. Conclusion： After infused through the vena caudalis, HUCBDSCs can home to the bone marrow, proliferating and differentiating there to reconstruct the impaired hematopoietic microenvironment.