尾静脉注射THP-1细胞构建急性髓系白血病NOD/SCID小鼠模型及其鉴定

目的:应用THP-1细胞构建NOD/SCID小鼠白血病模型。方法:将18只3-4周龄的雌性NOD/SCID小鼠,随机分为对照、模型A和模型B 3组(每组6只)。接种前连续2 d每只小鼠给予腹腔注射环磷酰胺2 mg/(kg·d),预处理后于24 h内接种细胞。模型组分别经尾静脉接种对数生长期THP-1细胞悬液1×10^7/只(A组)、5×106/只(B组),对照组鼠尾静脉注射等剂量生理盐水。观察一般情况,预处理前、接种细胞后7、14、21和28 d及处死时进行血常规检测、外周血白细胞分类,濒死前处死取材,组织切片病理检查。结果:模型组小鼠分别于接种细胞d 7和d 10开始出现竖毛、萎靡少动等现象,与对照组比较,模型A和B 2组小鼠体重于接种细胞21 d后明显下降(P<0.01),建模28 d后模型组白细胞数明显升高,差异有统计学意义(P<0.01),其中以接种1×10^7/只的模型A组最为显著。病理组织切片结果提示,模型组小鼠脾脏均可见白血病细胞弥漫性浸润。免疫组织化学结果提示,白血病细胞抗人CD13结果阳性,证实模型建立成功。结论:NOD/SCID小鼠经腹腔注射CTX预处理后,每只小鼠尾静脉注射THP-1细胞1×10^7或5×10^6个均可成功构建急性髓系白血病动物模型,符合急性髓系白血病的生物学特点,高浓度成瘤更快。

Construction and Identification of Acute Myeloid Leukemia NOD/SCID Mouse Model by Tail Vein Injection of THP-1 Cells

Objective:To construct NOD/SCID mouse leukemia model by using THP-1 cells.Methods:Eighteen female NOD/SCID mice aged 3 to 4 weeks were randomly divided into control group,model group A and model group B(6 in each group).Before inoculation,each mouse was intraperitoneally injected with cyclophosphamide 2 mg/(kg·d)for 2 d,and the mice in model groups were inoculated with cells within 24 h after pretreatment.The mice in model group were inoculated with THP-1 cell suspension in logarithmic growth phase by 1×10^7 cells/group(group A)and 5×10^6 cells/group(group B),the mice in the control group were injected with the same amount of normal saline in the tail vein.The general situation was observed,blood routine test and peripheral blood leukocyte classification were performed at 7,14,21,28 d of inoculation before the pre-treatment,and at the time sacrifice.Before dying,tissue of mice were collected and histological examination was performed.Results:Pilereation,droopiness and hypkinesia could be observed from d 7 and d 10 of inoculation cells in model group.Compared with the control group,the body weight of the mice in model group A and B decreased significantly after 21 days of inoculation(P<0.01),and the white blood cell counts increased significantly after 28 days of modeling(P<0.01).Among them,the above-mentioned presentation in inoculation of 1×10^7 group A was the most significant.Histopathological sections showed diffuse infiltration of leukemia cells in the spleen of the model group.The immunohistochemistry results indicated that the leukemia cells were positive for anti-human CD13,which confirmed the successful establishment of the model.Conclusion:After pretreatment with intraperitoneal injection of CTX in NOD/SCID mice,the injection of 1×10^7 or 5×10^6 THP-1 cells in tail vein of each mouse can successfully construct an acute myeloid leukemia animal model.The tumor formation is more much faster by injection of high concentration THP-1 cells.