人胚肺细胞系(MU-L1)生物学特性研究  

Biological Characteristics of Human Embryonic Lung Cell Line(MU-L1)

作　　者：田玲[1,3] 乔自林 杨迪 王家敏[1,2,4] 刘振斌 马忠仁[1,2,4] 马桂兰 王明明[3] TIAN Ling;QIAO Zi-lin;YANG Di;WANG Jia-min;LIU Zhen-bin;MA Zhong-ren;MA Gui-lan;WANG Ming-ming(Engineering Research Center of Key Technology and Industrialization of Cell-based Vaccine,Ministry of Education,Lanzhou 730030,China;Gansu Tech Innovation Center of Animal Cell,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,China;Life Science and Engineering College,Northwest Minzu University,Lanzhou 730030,China;Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030 China;Bioengineering Materials Engineering Research Center of Gansu Province,Lanzhou 730010,China;Department of Experiment&Teaching,Northwest Minzu University,Lanzhou 730030,China)

机构地区：[1]细胞基质疫苗关键技术与产业化教育部工程研究中心,甘肃兰州730030 [2]西北民族大学生物医学研究中心甘肃省动物细胞技术创新中心,甘肃兰州730030 [3]西北民族大学生命科学与工程学院,甘肃兰州730030 [4]西北民族大学生物医学研究中心生物工程与技术国家民委重点实验室,甘肃兰州730030 [5]甘肃省生物工程材料工程研究中心,甘肃兰州730010 [6]西北民族大学实验教学部,甘肃兰州730030

出　　处：《西北民族大学学报（自然科学版）》2023年第1期25-32,共8页Journal of Northwest Minzu University(Natural Science)

基　　金：西北民族大学研究生科研创新项目(Yxm2021083,Yxm2021079);中央高校基本科研业务费专项资金项目(31920220022)。

摘　　要：目的:对新建人胚肺细胞系(MU-L1)的生物学特性进行评价,以期建立可用于疫苗生产的人二倍体细胞株.方法:使用不同培养基及血清、不同血清浓度和传代比例培养MU-L1细胞,通过细胞形态、增殖速率和生长动力学等进行比较分析,筛选出较适宜的培养条件,用适宜的培养条件连续培养细胞至生命周期结束.传代过程中检查无菌、支原体和外源病毒因子,并进行染色体核型分析.结果:在常用培养基(MEM、DMEM、M199、DME/F12和PRMI1640)、胎牛血清和新生牛血清中MU-L1细胞均良好生长;按1∶4传代比例,使用含10%新生牛血清的MEM培养基培养,可培养至67代(世代),在生命周期的不同阶段经液氮冻存,复苏活率在91.56%~98.92%,生长良好.染色体核型为正常人二倍体核型,2n=46,无菌、支原体和外源病毒因子检查均为阴性.结论:MU-L1为正常人二倍体细胞系,具备作为病毒性疫苗基质进一步研究和生产应用的价值.Objective:In order to establish a human diploid cell line that can be used for vaccine production,we evaluated the biological characteristics of a newly established human embryonic lung cell line(MU-L1).Methods:To determine the most suitable culture conditions,MU-L1 cells were cultured with different medium and serum,different serum concentrations and passage ratios and the biological characteristics(cell morphology,proliferation rate and growth kinetics) were compared between different treatment groups.MU-L1 wascontinuously cultured under selected conditions until the end of their life cycle.In addition,sterility,mycoplasma,exogenous viral factors and karyotypes were examined during the passage stage.Results:MU-L1 grew well when cultured with common mediums(MEM,DMEM,M199,DME/F12 and PRMI1640),fetal bovine serum or newborn bovine serum.The treatment group cultured with MEM medium containing 10% newborn bovine serum and a passage ratio of 1:4 and can be cultivated to 67 generations.And its growth status was well after cryopreservation in liquid nitrogen with the recovery rate of 91.56-98.73%.Results of karyotype analysis showed that number of chromosomes was 2n=46,which was the same as the normal human diploid chromosomal status.Examinations of sterility,mycoplasma and exogenous viral factors all showed negative results.Conclusion:We provided a MU-L1 cell line with characteristics of normal human diploid cells line,which can be used for further research and applied as a potential material for viral vaccine production.

关 键 词：人胚肺细胞 培养条件 生物学特性 

分 类 号：R563[医药卫生—呼吸系统]