绿色荧光转基因大鼠模型的建立

目的随着干细胞研究的推进,大鼠干细胞的研究日趋迫切。本研究旨在为活体荧光影像系统、干细胞归巢、细胞移植体内示踪研究,提供绿色荧光蛋白EGFP转基因大鼠模型。方法通过显微注射方式获得EGFP转基因大鼠,采用活体荧光影像系统、激光共聚焦显微镜,对EGFP转基因大鼠各个组织的荧光表达水平进行比较;采用流式细胞术检测转基因大鼠血液和骨髓细胞、骨髓干细胞的荧光标记率,筛选骨髓干细胞高效标记绿色荧光的转基因大鼠。结果建立了心脏、肝脏、肌肉、肺、胰腺、脑、膀胱、胃、肾脏、肠和脾脏组织中,系统性表达EGFP的SD-TgN(ACT-EGFP-1)ZLFILAS转基因大鼠;流式细胞术检测表明,该品系血液细胞绿色荧光标记率为94.4%,骨髓干细胞绿色荧光标记率为97.8%。结论建立了多组织系统性高表达绿色荧光,骨髓干细胞荧光标记率高达95%以上的转基因大鼠,为影像分析,造血干细胞的归巢等研究提供了大鼠模型。

Establishment of Green Fluorescence Transgenic Rat

Object With the development of the stem cell, the study of rat stem cell became urgent. The purpose of this study was to generate EGFP transgenic rat models for fluorescence imaging, tracking of stem cells homing and cell transplantation. Methods The enhanced green fluorescent protein (EGFP) transgenic rats were obtained by microinjection and the expression of EGFP was determined under in vivo fluorescence imaging system and laser scanning confocal microscope. The expression of EGFP in blood cells, bone marrow cells and bone marrow stem cells was investigated with flow cytometry. Results The EGFP transgenic rat SD-TgN(CAG-EGFP-1)ZLFILAS was established. EGFP was systematically expressed in tissues of heart, liver, muscle, lung, pancreas, brain, bladder, stomach, kidney, intestines and spleen, and EGFP labeling ratios in blood cells and bone marrow stem cells were 94.4% and 97.8% respectively. Conclusions A systematic EGFP transgenic rat line was generated which showed EGFP labeling ratios higher than 95% in bone marrow stem cells. This transgenic rat may be a useful rat tool for fluorescence imaging and stem cells tracking.