1. Genotype instability during long-term subculture of lymphoblastoid cell lines.
- Author
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Oh, Ji Hee, Kim, Young Jin, Moon, Sanghoon, Nam, Hye-Young, Jeon, Jae-Pil, Ho Lee, Jong, Lee, Jong-Young, and Cho, Yoon Shin
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LYMPHOBLASTOID cell lines ,EPSTEIN-Barr virus ,DNA ,HUMAN chromosomes ,BLOOD cells ,SINGLE nucleotide polymorphisms ,HETEROZYGOSITY - Abstract
Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines (LCLs) promise to address the challenge posed by the limited availability of primary cells needed as a source of genomic DNA for genetic studies. However, the genetic stability of LCLs following prolonged culture has never been rigorously investigated. To evaluate genotypic errors caused by EBV integration into human chromosomes, we isolated genomic DNA from human peripheral blood mononuclear cells and LCLs collected from 20 individuals and genotyped the DNA samples using the Affymetrix 500K SNP array set. Genotype concordance measurements between two sources of DNA from the same individual indicated that genotypic discordance is negligible in early-passage LCLs (<20 passages) but substantial in late-passage LCLs (>50 passages). Analysis of concordance on a chromosome-by-chromosome basis identified genomic regions with a high frequency of genotypic errors resulting from the loss of heterozygosity observed in late-passage LCLs. Our findings suggest that, although LCLs harvested during early stages of propagation are a reliable source of genomic DNA for genetic studies, investigations that involve genotyping of the entire genome should not use DNA from late-passage LCLs. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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