【摘要】 目的建立唐氏综合征患者永生细胞系,为唐氏综合征的基因诊断提供标准品。方法采集唐氏综合征患者外周血,采用 EB 病毒转化外周血 B 淋巴细胞和加环孢霉素 A 抑制 T 淋巴细胞的技术,建立永生淋巴母细胞系。用细胞遗传学方法、实时荧光定量 PCR 及短串联重复序列(STR)多态性分析技术检测其建系前后的遗传特性及稳定性。结果建系成功,经过传代、冻存和复苏,建系后第10代染色体核型和 DNA 标志未发生改变。建系前后染色体核型均为47,XX,+21。多重荧光相对定量 PCR 鉴定,唐氏综合征患者外周血原代细胞和转化后第10代细胞株的△Ct 值均在-2.55±3s 范围内,与正常对照相比△Ct 值范围无重叠。21号染色体上的 STR 基因座 D21S11多态性分析表明,唐氏综合征患者外周血原代细胞和转化后第10代细胞株均有同样的3种基因型。结论染色体核型和 DNA 分析表明,唐氏综合征患者永生细胞系建立前后遗传特点一致,且稳定,可用作基因诊断试剂盒产品质量检测、质量控制的标准品。更多还原

【Abstract】 Objective Establishing the immortalized lymphoblastoid cell line of Down syndrome,in order to supply the standard for genetic diagnosis of Down syndrome.Methods The peripheral blood sample from a patient of Down syndrome was collected.The immortal lymphoblastoid cell line was established by Epstein-Barr virus transformation of B cells and addition of cyclosporine A to inhibit the activity of T cells. Genetic stability of the cell line was checked by karyotype analysis,multiplex real time PCR and short tandem repeat(STR)polymorphism analysis.Results The immortalized cell line was established and preserved.The freeze preserved cells could be resuscitated successfully.The chromosome karyotype was 47, XX,+21 in both pre-and post-immortal lymphoblastoid cell line.Multiplex real time PCR tests indicated no overlapping sphere of delta Ct values between the immortalized cell line and the normal control. Polymorphism of STR locus 1321S11 showed same 3 genotypes in the primary cells and the immortalized cell line.Conclusions Genetic integrity of the trisomy 21 immortal lymphoblastoid cell line is stable.The cell line could be used as standard in quality control for Down syndrome genetic diagnosis.更多还原