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Long-term Culture of EBV-induced Human Lymphoblastoid Cell Lines Reveals Chromosomal Instability.
Journal of Histochemistry & Cytochemistry ( IF 3.2 ) Pub Date : 2020-02-28 , DOI: 10.1369/0022155420910113
Marianne Volleth 1 , Martin Zenker 1 , Ivana Joksic 2 , Thomas Liehr 3
Affiliation  

To preserve material for future genetic studies, human B-lymphocytes from whole blood samples are routinely transformed into lymphoblastoid cell lines (LCLs) by in vitro infection with Epstein-Barr virus. To determine the rate and frequency of chromosomal changes during long-term culture, we established 10 LCLs (from eight individuals). Before transformation, these cases showed a normal karyotype (three cases), a small supernumerary marker chromosome (three cases), or an aberrant karyotype (four cases). Chromosome analyses were performed at 8-week intervals over a period of at least 1 year, up to 3 years. Surprisingly, we demonstrate that chromosomal instability is the rule, rather than the exception, during long-term culture of LCLs. The most commonly observed acquired clonal aberration was trisomy 12, which emerged in all cell lines within 21 to 49 weeks after infection. Telomeric fusions indicating telomere shortening were found after ~21 weeks. After 1 year of cultivation, the proportion of cells with the original karyotype decreased to ≤10% in 7 of the 10 cell lines. To preserve cells with aberrant genomes, we conclude the cultivation time of LCLs must be restricted to the absolute minimum time required.

中文翻译:

EBV诱导的人淋巴母细胞细胞系的长期培养揭示了染色体的不稳定性。

为了保存用于将来的遗传研究的材料,通过体外感染爱泼斯坦-巴尔病毒将全血样品中的人B淋巴细胞常规转化为淋巴母细胞样细胞系(LCL)。为了确定长期培养过程中染色体变化的速率和频率,我们建立了10个LCL(来自8个个体)。在转化之前,这些病例显示出正常的核型(3例),小的数字标记染色体(3例)或异常的核型(4例)。在至少1年至3年的时间内,每隔8周进行一次染色体分析。出乎意料的是,我们证明了在LCL的长期培养过程中,染色体不稳定是规则,而非例外。最常见的获得性克隆畸变是三体性12,感染后21至49周内所有细胞系中均出现了这种病毒。约21周后发现表明端粒缩短的端粒融合体。培养1年后,在10个细胞系中的7个中,具有原始核型的细胞比例下降到≤10%。为了保存具有异常基因组的细胞,我们得出结论,LCL的培养时间必须限制在所需的绝对最短时间。
更新日期:2020-04-21
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