摘要
白血病研究中常需要在细胞中进行慢病毒介导的基因过表达或者敲降,但慢病毒生产方法存在成本高、对悬浮细胞感染效率低等问题。为解决这些问题,通过比较利用线性聚乙烯亚胺(linear polyethylenimine,LPEI)和脂质体两种转染方法包装成的慢病毒对白血病细胞的感染能力,以及感染后对细胞生物学特性的影响,确定不同转染方法在白血病实验研究中的有效性和安全性。在相同条件下利用LPEI和脂质体产生慢病毒颗粒,比较发现用LPEI转染法包装的慢病毒滴度略高。根据病毒感染复数和慢病毒滴度分别感染3种白血病细胞系(K562、HL60和HEL),两种方法间感染白血病细胞系效率无差别。在感染不同细胞系时两种不同转染方法对细胞增殖略有影响,但对其他生物学特性比如细胞诱导分化和克隆形成能力的影响是一致的。因此在白血病相关研究中,转染试剂LPEI包装慢病毒方法可以成为更加经济实用的选择。
Gene overexpression and knock down mediated by lentivirus is often used in leukemia research. There are some problems, such as high cost and low efficiency in suspension cell infection. In order to identify the effectiveness and safety of the lentivirus packaged with different transfection method in leukemia research, solving the problems of low infection efficiency and high cost during the lentivirus production, we packaged lentiviruses with linear polyethylenimine (LPEI) and liposomes, and then compared the infection ability and the effects on biological characteristics of leukemia cell lines of lentiviruses produced with these two methods. Using LPEI or liposomes to generate lentiviral particles, respectively, our studies showed that the titer of lentivirus packaged with LPEI was slightly higher than that packaged with liposomes under the same packaging conditions. Three leukemia cell lines (K562, HL60 and HEL) were infected with the same amount lentiviral particles according to multiplicity of cellular infection (MOI) and lentiviral titer, and there was no difference in infection efficiency between the two groups. The cell proliferation of leukemic cells infected with lentivirus produced with LPEI was slightly slower than that in liposome group, the effects on induced differentiation and colony forming ability of these two methods were similar. Therefore, using lentivirus produced with LPEI transfection method can be an economical and practical choice in experimental studies in leukemia.
作者
张明英
袁佳佳
张晓茹
邢文
白洁
周圆
ZHANG Mingying;YUAN Jiajia;ZHANG Xiaoru;XING Wen;BAI Jie;ZHOU Yuan(State Key Laboratory of Experimental Hematology,Institute of Hematology and Blood Diseases Hospital,Chinese Academy of Medical Science&Peking Union Medical College,Tianjin 300020,China;The Second Hospital of Tianjin Medical University,Tianjin 300020,China)
出处
《生物技术进展》
2019年第3期262-270,共9页
Current Biotechnology
基金
国家自然科学基金项目(81170228)
中国医学科学院医学与健康科技创新工程项目(2016-I2M-2-001)
天津市应用基础研究重点项目(16JCZDJC34500)资助
