脂质体介导SV40LT基因转染构建羊水染色体核型分析质控细胞系研究  

Establishment of cell lines for quality control of prenatal genetic diagnosis by SV40LT gene transfection

作　　者：翁炳焕[1] 徐威[2,3] 苏岚 沈敏 李蓉[3] 虞晓鹏 李兰娟[2,3] WENG Binghuan;XU Wei;SU Lan;SHEN Min;LI Rong;XU Xiaopeng;LI Lanjuan(Key Laboratory of Reproductive Genetics of the Ministry of Education,Women's Hospital,Zhejiang University School of Medicine,Hangzhou 310006,China;State Key Laboratory for Diagnosis and Treatment of Infectious Diseases,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases,the First Affiliated Hospital,Zhejiang University School of Medicine,Hangzhou 310003,China;Shulan (Hangzhou )Hospital,Hangzhou 310004,China)

机构地区：[1]浙江大学医学院附属妇产科医院生殖遗传教育部重点实验室,浙江杭州310006 [2]浙江大学医学院附属第一医院传染病诊治国家重点实验室感染性疾病诊治协同创新中心,浙江杭州310003 [3]树兰(杭州)医院,浙江杭州310004

出　　处：《浙江大学学报（医学版）》2018年第5期520-524,共5页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金(81671467);传染病诊治国家重点实验室开放基金(2017KF06)

摘　　要：目的:研究羊水染色体核型分析质控细胞系的构建方法。方法:以T4DNA连接经BamHⅠ单酶切的SV40LTag-pcDNA和pcDNA3.1 (-) DNA,重组SV40LTag-pcDNA3.1(-)克隆,以脂质体介导法将重组克隆转染至染色体结构异常的羊水细胞,以G418筛选阳性克隆,观察细胞系的传代生长特性及其作为染色体核型分析质量评估的可行性。结果:构建了染色体核型为46,XY,t(8; 19)(q24.3;q13.1)的羊水细胞系,传至第15代的细胞系经染色体核型分析,其核型与原代细胞一致。结论:染色体结构异常的羊水细胞转染SV40LT基因后可转化为无限扩增且染色体核型稳定的细胞系,从而制成羊水细胞染色体核型分析的质控细胞系。Objective:To establish a cell lines for quality control of prenatal genetic diagnosis.Methods:The recombined SV40LTag-pcDNA3.1(-) vector was constructed and transfected by lipidosome into human amniotic fluid cells with common aneuploidy.Positive clones were screened by G418,and the immortality of transfected cell line was identified.Results:Cell line with karyotype of 46,XY,t(8;19)(q24.3;q13.1) from primary amniotic fluid cells was established.Karyotype analytical results indicated that the cell line at its 15 th generation maintained the same karyotype of its primary cell.Conclusion:Gene SV40 LT can lead to immortality of amniotic fluid cells,which contributes to preparing cell lines for internal and external quality control in prenatal genetic diagnosis.

关 键 词：猴病毒40/免疫学 猴病毒40/遗传学 脂质体 转染 羊水/细胞学 遗传载体 质量控制 细胞 培养的 

分 类 号：R394[医药卫生—医学遗传学] R722.11[医药卫生—基础医学]