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

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.