绿色荧光蛋白转基因小鼠骨髓间充质干细胞干预重症急性胰腺炎大鼠后在多脏器的分布

背景：利用绿色荧光蛋白转基因小鼠间充质干细胞自身携带荧光性的特点,干预重症急性胰腺炎大鼠后,便于动物体内跟踪观察。目的：观察绿色荧光蛋白转基因小鼠骨髓间充质干细胞在重症胰腺炎大鼠体内各脏器的分布情况。方法：直接贴壁法分离培养绿色荧光蛋白转基因小鼠骨髓间充质干细胞,90%融合后消化传代扩增。传至第3代后行CD29＋、CD90＋、CD34-、CD45-细胞免疫表型鉴定。显微镜下逆行胰胆管注射5%牛黄胆酸钠制造SD大鼠重症急性胰腺炎模型。1h后,按2×106/只尾静脉注入重症急性胰腺炎SD大鼠体内。分别于6,12,24h取肝、肾、脑、肺、胰、肠脏器送普通病理检查,观察胰腺病理变化及骨髓间充质干细胞干预重症急性胰腺炎SD大鼠后在各脏器干细胞分布情况及灰度值测定。结果与结论：胰腺破坏随时间延长而加强,24h破坏最严重;GFP小鼠CD29阳性细胞91.1%,CD90阳性细胞93.5%,CD34阳性细胞0.82%,CD45阳性细胞2.22%;注射干细胞SD大鼠各脏器均有绿色荧光出现,并随时间增长而增强;在肾脏组织中灰度值最高,脑组织最少。提示骨髓间充质干细胞干预重症急性胰腺炎大鼠后能在各脏器稳定分布。

Distribution of green fluorescent protein transgenic mouse bone marrow mesenchymal stem cells in various organs of rats with severe acute pancreatitis

BACKGROUND：Green fluorescent protein transgenic mouse bone marrow mesenchymal stem cells are transplanted into rats with severe acute pancreatitis for easy in vitro tracing in animals because of the characteristic of carrying fluorescence.OBJECTIVE：To investigate the distribution of green fluorescent protein transgenic mouse bone marrow mesenchymal stem cells in various organs of rats with severe acute pancreatitis.METHODS：Green fluorescent protein transgenic mouse bone marrow mesenchymal stem cells were isolated directly by adherence.After reaching 90% confluency,cells were digested,passaged,and proliferated.After three passages,cell immunophenotype of CD29＋,CD90＋,CD34-,CD45-were determined.Sprague-Dawley rat model of severe acute pancreatitis were established by retrograde injection of 5% sodium taurocholate into the pancreatic duct under the microscope.1 hour later,cells at a density of 2×106/rat were injected into rats with severe acute pancreatitis via tail vein.At 6,12,24 hours,the liver,kidney,brain,lung,pancreas,and intestine were harvested for pathological examination.The pathological changes of the pancreas and the distribution of green fluorescent protein transgenic mouse bone marrow mesenchymal stem cells in various organs of rats with severe acute pancreatitis as well as gray scale values were determined.RESULTS AND CONCLUSION：The pancreas was more seriously destroyed with the time and it was most seriously destroyed at 24 hours.91.1% of green fluorescent protein transgenic mouse mesenchymal stem cells were positive for CD29,93.5% positive for CD90,0.82% positive for CD34,and 2.22% positive for CD45.After injection of stem cells,green fluorescence was observed in each organ and enhanced with time.The gray scale of green fluorescence was highest in the kidney tissue and lowest in the brain tissue.These findings suggest that the transplanted bone marrow mesenchymal stem cells can be stably distributed in each organ of rats with severe acute pancreatitis.