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)

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.