应用MEC-1与HG3细胞株建立慢性淋巴细胞白血病BALB/c裸鼠皮下移植瘤的模型  

Establishment of Human Chronic Lymphocytic Leukemia Model in the BALB/c Nude Mice by Using MEC-1 and HG3 Cell Lines

作　　者：付春玲[1,2,3] 龚艳青[1,2,3] 万艳[1,2,3] 徐开林[1,2,3] 

机构地区：[1]徐州医学院 [2]徐州医学院附属医院干细胞实验室 [3]江苏省血液病研究所,江苏徐州221002

出　　处：《中国实验血液学杂志》2016年第6期1644-1648,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金青年基金(81400127);中国博士后科学基金(2015M571818)

摘　　要：目的：通过对免疫缺陷鼠（BALB/c）不同部位、不同密度接种人慢性淋巴细胞白血病（CLL）细胞,探索裸鼠皮下移植瘤模型建立的条件。方法：通过慢病毒系统建立稳定表达绿色荧光蛋白（GFP）的人CLL细胞（MEC-1-GFP与HG3-GFP细胞）。首先将MEC-1-GFP细胞（5×10^7/ml）分别接种于裸鼠的前肢腋下、腹部及后肢腋下,观察不同接种部位对CLL细胞成瘤率的影响;其次将同等密度的MEC-1-GFP、HG3-GFP细胞（5×10^7/ml）分别接种于裸鼠的前肢腋下部位,观察两种细胞的成瘤时间及成瘤率。再次以同样接种方法,将不同密度的MEC-1-GFP与HG3-GFP细胞（1×10^7/ml与5×10^7/ml）分别接种于裸鼠的前肢腋下,观察不同接种密度的细胞成瘤时间及成瘤率。观察5周后,通过眼球取血获得外周静脉血,经EDTA抗凝及溶血素处理后,用流式细胞术检测GFP阳性的CLL细胞;同时,将荷瘤鼠处死,剥离肿瘤组织并制备冰冻切片,进行组织病理学检查。结果：成功获得稳定表达标记蛋白GFP的MEC-1与HG3细胞株;接种后裸鼠的前肢腋下、腹部及后肢腋下都形成了皮下移植瘤,且前肢腋下更有利于移植瘤的形成;接种密度为5×10^7/ml的MEC-1-GFP和HG3-GFP细胞均可形成皮下移植瘤,且5周后的成瘤率均为80%;接种密度分别为1×10^7/ml与5×10^7/ml MEC-1-GFP与HG3-GFP细胞均可有效地形成皮下移植瘤。流式细胞术检测显示,2组荷瘤鼠外周血中均未见GFP阳性的MEC和HG3细胞,组织病理检查表明CLL细胞向腹腔转移。结论：利用CLL细胞株MEC-1-GFP和HG3-GFP建立了CLL裸鼠皮下移植瘤模型,为进一步研究CLL的发病机制提供了实验工具。Objective：To explore the the optimal condition for establishing immune deficiency mouse（BALB/c）model with CLL via subcutaneous inoculation of human chronic lymphocytic leukemia（CLL） cells at different inoculative locations and different cell concentrations.Methods：Firstly,Two CLL cell lines（MEC-1-GFP and HG3-GFP） with the green fluorescent protein（GFP） were established by lentivirus system respectively,and then the MEC-LGFP cells（5 X 10 /ml） were inoculated into forelimb,hindlimb and abdomen to observe the tumorigenesis.Secondly,the MEC-1-GFP and HG3-GFP cells with same density（5 × 10-7 /ml） were inoculated into forelimb to compare the time and rate of tumor formation.Thirdly,the MEC-1-GFP cells（1×10^7ml） and HG3-GFP cells（5×10^7ml） were inoculated into forelimb to compare the time and rate of tumor formation at different inoculative density.After observation for 5 weeks,the peripheral blood was collected and treated with EDTA and erythrocytolysin,then the of GFP positive cells were detected by flow cytomety.Meanwhile,the tumor- bearing mice were killed,and the tumors were isolated and cut into slices for histopathological examination.Results：MEC-L-GFP and HG3-GFP cell lines were successfully established,and after inocutation of MEC-1-GFP cells with 5 × 10-7/ml the xenograt tumors were formated in forelimb,hindlimb and abdomen of mice,especially in the forelimb with a higher tumorigenic rate.In addition,the inoculation of same density of MEC-1-GFP and HG3-GFP cells（5×10^7ml） also resulted in xenograft in forelimb,and the tumorigenic rate reached to 80%after 5 weeks.Moreover,the inocutation of MEC-L-GFP and HG3-GFP cells with1×10^7and 5×10^7/ml respectively also effectively resulted to xenograft tumor in forelimb.The flow cytometry showed that there was no MEC-1-GFP and HG3-GFP cells in peripheral blood,while histopathological examination demonstrated CLL cell metastasis towards peritoneal cavity.Conclusion：The BALB/c nude mouse model is successfully established b

关 键 词：慢性淋巴细胞白血病 MEC-1-GFP、HG3-GFP细胞 BALB/C裸鼠模型 皮下移植瘤 

分 类 号：R733.72[医药卫生—肿瘤]