应用新型免疫缺陷型NCG小鼠建立患者来源急性T淋巴细胞白血病模型的研究  

Establishment of a Patient-Derived T-Cell Acute Lymphoblastic Leukemia Xenograft Model in Novel Immunodeficient NCG Mice

作　　者：姜鹏君[1] 代兴斌[1] 孔祥图[1] 徐祖琼[1] 于慧[1] 庞洁[1] 夏雯[1] 于菊华 朱光荣[1] 田芳 朱学军[1] JIANG Peng-Jun;DAI Xing-Bin;KONG Xiang-Tu;XU Zu-Qiong;YU Hui;PANG Jie;XIA Wen;YU Ju-Hua;ZHU Guang-Rong;TIAN Fang;ZHU Xue-Jun(Department of Hematology,The Affiliated Hospital of Nanjing University of Chinese Medicine,Jiangsu Province Hospital of Chinese Medicine,Nanjing 210029,Jiangsu Province,China;Central Laboratory,The Affiliated Hospital of Nanjing University of Chinese Medicine,Jiangsu Province Hospital of Chinese Medicine,Nanjing 210029,Jiangsu Province,China)

机构地区：[1]南京中医药大学附属医院,江苏省中医院血液科,江苏南京210029 [2]南京中医药大学附属医院,江苏省中医院中心实验室,江苏南京210029

出　　处：《中国实验血液学杂志》2023年第2期311-318,共8页Journal of Experimental Hematology

基　　金：国家自然科学基金(NO.81673771);江苏省社会发展-临床前沿技术项目(BE2016809);南京市科技发展计划项目(201503011);全国中医药创新骨干人才培训项目(2019-128);江苏省中医院院级课题(Y17014)。

摘　　要：目的:将患者来源的急性T淋巴细胞白血病(T-ALL)细胞接种于NCG小鼠,建立稳定的人类T-ALL白血病动物模型。方法:分离初诊T-ALL患者骨髓来源的白血病细胞,鉴定后经尾静脉接种于NCG小鼠。定期采用流式细胞术检测小鼠外周血中hCD45阳性的细胞比例,通过病理及免疫组化检测小鼠骨髓、肝脏、脾脏等脏器组织白血病细胞浸润情况。第一代小鼠模型建立成功后,取脾脏细胞接种于第二代小鼠,第二代小鼠模型建立成功后取脾脏细胞进一步接种于第三代小鼠,定期采用流式细胞术监测各组小鼠外周血白血病细胞生长情况,以评估此T-ALL白血病动物模型的稳定性。结果:第一代小鼠在接种d 10可在外周血中检测到hCD45阳性的白血病细胞,其后比例逐渐增高,平均第6-7周小鼠出现精神萎靡,小鼠外周血及骨髓涂片可见大量T淋巴细胞白血病细胞。小鼠脾脏明显肿大,免疫组织化学检测结果显示hCD3阳性的白血病细胞广泛浸润骨髓、肝脏、脾脏等。第二代、第三代小鼠能稳定发生白血病,平均生存期为4-5周。结论:采用T-ALL患者骨髓来源的白血病细胞,经尾静脉接种于NCG小鼠,可以成功构建患者来源的肿瘤细胞异种移植(PDTX)模型。Objective:The leukemia cells from patients with T-cell acute lymphoblastic leukemia(T-ALL)were inoculated into NCG mice to establish a stable human T-ALL leukemia animal model.Methods:Leukemia cells from bone marrow of newly diagnosed T-ALL patients were isolated,and the leukemia cells were inoculated into NCG mice via tail vein.The proportion of hCD45 positive cells in peripheral blood of the mice was detected regularly by flow cytometry,and the infiltration of leukemia cells in bone marrow,liver,spleen and other organs of the mice was detected by pathology and immunohistochemistry.After the first generation mice model was successfully established,the spleen cells from the first generation mice were inoculated into the second generation mice,and after the second generation mice model was successfully established,the spleen cells from the second generation mice were further inoculated into the third generation mice,and the growth of leukemia cells in peripheral blood of the mice in each group was monitored by regular flow cytometry to evaluate the stability of this T-ALL leukemia animal model.Results:On the 10th day after inoculation,hCD45^(+)leukemia cells could be successfully detected in the peripheral blood of the first generation mice,and the proportion of these cells was gradually increased.On average,the mice appeared listless 6 or 7 weeks after inoculation,and a large number of T lymphocyte leukemia cells were found in the peripheral blood and bone marrow smear of the mice.The spleen of the mice was obviously enlarged,and immunohistochemical examination showed that hCD3^(+)leukemia cells infiltrated into bone marrow,liver and spleen extensively.The second and third generation mice could stably develop leukemia,and the average survival time was 4-5 weeks.Conclusion:Inoculating leukemia cells from bone marrow of patients with T-ALL into NCG mice via tail vein can successfully construct a patient-derived tumor xenografts(PDTX)model.

关 键 词：急性T淋巴细胞白血病 异种移植 免疫缺陷 NCG小鼠 动物模型 

分 类 号：R-332[医药卫生] R733.7