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首页> 外文期刊>Biotechnology Progress >Chromosome instability in spodoptera frugiperda Sf-9 cell line
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Chromosome instability in spodoptera frugiperda Sf-9 cell line

机译：草地贪夜蛾Sf-9细胞系的染色体不稳定

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摘要

Homogeneous cell lines are essentail in industry and research if reliable and reproducible data are to be obtained.The Spodoptera frugiperda (Sf-9) cell line routinely used for the production of recombinant proteins was found to be heterogeneous,containing a mixture of diploid and tetraploid cells.Using dilution-cloning techniques,diploid and tetraploid subpopulations were isolated from a Sf-9 parentalcell line,and their cytogenetic state was monitored using Vinblastine toarrest cells in mitosis.Flow cytometry was sued to obtaina snapshot of the predominant subpopulations present to verify the karyological results.The rate at which clonal populations digress into the heterogeneous state was found to be more rapid for the diploid subpopulation,with the emergence of tetraploid cells after only 11 passages,than for tetraploid subpopulation,where diploid clones appeared after 18 passages.The chromosomes in both diploid and tetraploid subpopulations as well as the parental cell line were found to spontaneously fragment during growth and expansion processes giving rise tovariable chromosome numbers.NDA analysis of cell lines obtained from laboratories worldwide have shown that the Sf-9 cell line used for the production of many recombiant proteins is cytologically unstable,leading to varying degrees of polyploidal state depending on its culture history and supplier.
机译：如果要获得可靠和可重复的数据，同质细胞系在工业和研究中尤为重要。发现通常用于生产重组蛋白的贪食夜蛾（Sf-9）细胞系是异质的，其中包含二倍体和四倍体的混合物。使用稀释克隆技术，从Sf-9亲本细胞系中分离出二倍体和四倍体亚群，并使用长春碱近邻细胞在有丝分裂中监测它们的细胞遗传状态。使用流式细胞术获得当前主要亚群的快照以验证其存在性。二倍体亚群的克隆种群进入异质状态的速度更快，四倍体亚群仅在11代后出现，四倍体亚群则在18代后出现二倍体克隆。二倍体和四倍体亚群的染色体以及亲本细胞系均为并在生长和扩增过程中自发断裂，从而产生可变的染色体数。从全球实验室获得的细胞系的NDA分析表明，用于生产许多重组蛋白的Sf-9细胞系在细胞学上不稳定，导致不同程度的多倍体状态取决于其培养历史和供应商。

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