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目的探讨含绿色荧光蛋白慢病毒(LV-GFP)和荧光染料四甲基-吲哚羧花青-高氯酸盐(CM-Di I)两种标记干细胞的示踪技术在人脐带间充质干细胞(HU-MSCs)治疗子痫前期(PE)大鼠研究中的应用。方法采用酶消化法分离培养HU-MSCs,分别以LV-GFP和CM-Di I标记HU-MSCs。建立脂多糖(LPS)诱导的PE大鼠模型,移植HU-MSCs后分别通过荧光显微镜和活体成像技术示踪HU-MSCs的分布。结果荧光显微镜下见LV-GFP标记的HU-MSCs分布于大鼠胎盘组织和肾脏组织中;胎鼠肾脏、肝脏、心脏和肺脏组织中未见分布。活体成像技术显示CM-Di I标记的HU-MSCs分布于大鼠胸腹部脏器及胎盘,而脐带及胎鼠内未见分布。结论 LV-GFP标记后荧光显微镜可显示最终HU-MSCs归巢并定位准确,CM-Di I标记后活体成像技术则可以观察活体内HU-MSCs分布变化。
Objective To investigate the effect of tracing technique of green fluorescent protein lentivirus (LV-GFP) and fluorescent dye tetramethylindole carboxylase-perchlorate (CM-Di I) on human umbilical cord mesenchymal Application of stem cells (HU-MSCs) in the treatment of preeclampsia (PE) rats. Methods HU-MSCs were isolated and cultured by enzymatic digestion. HU-MSCs were labeled with LV-GFP and CM-Di I, respectively. The rat model induced by lipopolysaccharide (LPS) was established. After the HU-MSCs were transplanted, the distribution of HU-MSCs was observed by fluorescence microscopy and in vivo imaging respectively. Results Fluorescence microscopy showed that LV-GFP-labeled HU-MSCs distributed in rat placenta tissue and kidney tissue. No distribution was observed in fetal kidney, liver, heart and lung tissues. In Vivo imaging, CM-Di I-labeled HU-MSCs were distributed in the organs and placenta of the thorax and abdomen in rats, but not in the umbilical cord and fetal rats. Conclusion LV-GFP labeled post-fluorescence microscopy showed that the final HU-MSCs homing and localization accuracy, CM-Di I labeled live imaging technology can be observed in vivo HU-MSCs distribution changes.