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目的探讨用非肥胖型糖尿病/严重联合免疫缺陷(NOD/SCID)小鼠和经流式细胞仪分选出的KG1a中CD34+CD38-的干细胞亚群建立白血病干细胞(LSCs)动物模型,为靶向治疗LSCs的药物筛选奠定体内实验基础。方法 18只雌性NOD/SCID小鼠,体质量18～20 g,鼠龄6～8周龄。实验组12只,应用流式细胞仪分选KG1a细胞中具有LSCs特性的CD34+CD38-亚群,以2×106个/只尾静脉注射经全身X射线照射2Gy的NOD/SCID小鼠;正常对照组6只,只注射磷酸盐缓冲溶液。观察两组小鼠的一般情况和白血病发生情况,应用形态学、组织病理检查、流式细胞术、骨髓染色体检查等检测实验组小鼠的外周血、骨髓、肝脏、脾脏的白血病细胞标志。结果接种2周后实验组小鼠外周血可见白血病细胞,接种30 d实验组小鼠的白血病发病率为100%,无自发缓解;外周血、骨髓、肝、脾中均可发现大量的白血病细胞浸润;实验组小鼠骨髓细胞中CD13抗原阳性率15.47%～23.66%,并可见KG1a细胞的核型特征。结论尾静脉接种流式细胞仪分选后的CD34+CD38-KG1a细胞于全身亚致死量X射线照射后的NOD/SCID小鼠,能成功建成全身播散的白血病模型,为进一步研究LSCs的靶向治疗药物奠定实验基础。
OBJECTIVE: To establish an animal model of leukemia stem cells (LSCs) using non-obese diabetic / severe combined immunodeficiency (NOD / SCID) mice and CD34 + CD38- stem cell subsets of KG1a sorted by flow cytometry Drug therapy for the treatment of LSCs laid the foundation for in vivo experiments. Methods Eighteen female NOD / SCID mice, weighing 18-20 g, were aged 6-8 weeks. In the experimental group, 12 CD34 + CD38 subpopulations of KG1a cells with LSCs were sorted by flow cytometry and 2Gy of NOD / SCID mice were irradiated with 2 × 106 / Control group of 6, only injection of phosphate buffer solution. The general situation of the two groups of mice and the occurrence of leukemia were observed. The leukemic cell markers in peripheral blood, bone marrow, liver and spleen of experimental mice were detected by morphological, histopathological examination, flow cytometry and bone marrow chromosome examination. Results The leukemia cells were found in the peripheral blood of mice in the experimental group two weeks after inoculation. The incidence of leukemia was 100% in the mice on the 30th day after inoculation, without spontaneous remission. A large number of leukemia cells were found in peripheral blood, bone marrow, liver and spleen Infiltration; experimental group mice bone marrow CD13 antigen positive rate of 15.47% ~ 23.66%, and can be seen KG1a cell karyotype characteristics. CONCLUSIONS: After the tail vein is infused with CD34 + CD38-KG1a cells sorted by flow cytometry, whole-body lymphocyte subsets can be successfully established in NOD / SCID mice after systemic sublethal X-ray irradiation. In order to further study the target of LSCs To lay the foundation for experimental treatment of drugs.