建立非肥胖型糖尿病/重症联合免疫缺陷(NOD/SCID)小鼠人白血病模型的实验研究

The establishment of nonobese diabetic-severe combined immune deficient(NOD/SCID)mice model inoculated with GFP-encoded K562 leukemia cells

目的:建立绿色荧光蛋白(GFP)标记的人白血病细胞株K562,并以此移植非肥胖型糖尿病/重症联合免疫缺陷(NOD/SCID)小鼠,建立白血病研究的新动物模型。方法:用含GFP的逆病毒载体将标记基因GFP转入人白血病细胞株K562。NOD/SCID小鼠经2.5Gy的γ射线照射后,从尾静脉注射0.5×106个K562-GFP细胞(n=3;第1组);或1×106个K562-GFP细胞(n=3;第2组);或生理盐水作为对照组(n=2)。移植后第6周用流式细胞术和PCR检测受鼠体内白血病细胞。结果:外源性GFP基因在K562细胞中稳定表达。移植后6周FCM检测,受鼠骨髓中的人源细胞比例均数分别为12.3%(第1组)和22.6%(第2组),并且外周血和脾脏也可检测白血病细胞。结论:用GFP标记的K562细胞移植NOD/SCID小鼠,成功建立了白血病动物模型。

Objective：To establish the nonobese diabetic-severe combined immune deficient（NOD/SCID） mice model inoculated with human leukemia cell line K562 cells,which encoded a marker gene-green fluorescence protein. Methods：GFP-encoded K562 cells were obtained by the infection of K562 cells with recombinant retrovirus encoded GFP. After total body irradiation with γ-ray of 2.5Gy,NOD/SCID mice were intravenously transplanted with 5 ×10^5 K562-GFP cells （Group 1 ,n = 3）,or 1 ×10^6 K562-GFP cells （Group 2, n = 3）, or normal saline （Control, n = 2）. Six weeks after transplantation, the bone marrow, peripheral blood and spleen of recipient mice were assayed for the presence of human cells by flow cytometry and PCR. Results：Six weeks posttransplatation,the presence of human leukemia cells was found in the peripheral blood,bone marrow and spleen of recipient mice. The mean percentages of human cells in the bone marrow of recipient mice of Group 1 and Group 2 were 12.3% and 22.6% ,respectively. Conclusion：human leukemia model inoculated with GFP-encoded k562 cells was successfully established,which could be employed in the future research.