Genotype instability during long-term subculture of lymphoblastoid cell lines

Ji Hee Oh, Young Jin Kim, Sanghoon Moon, Hye Young Nam, Jae Pil Jeon, Jong Ho Lee, Jong Young Lee, Yoon Shin Cho

Research output: Contribution to journalArticle

19 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)16-20
Number of pages5
JournalJournal of human genetics
Volume58
Issue number1
DOIs
Publication statusPublished - 2013 Jan 1

Fingerprint

Genotype
Cell Line
DNA
Human Herpesvirus 4
Chromosomes
Virus Integration
Transformed Cell Line
Loss of Heterozygosity
Human Chromosomes
Single Nucleotide Polymorphism
Blood Cells
Genome

All Science Journal Classification (ASJC) codes

  • Genetics
  • Genetics(clinical)

Cite this

Oh, Ji Hee ; Kim, Young Jin ; Moon, Sanghoon ; Nam, Hye Young ; Jeon, Jae Pil ; Lee, Jong Ho ; Lee, Jong Young ; Cho, Yoon Shin. / Genotype instability during long-term subculture of lymphoblastoid cell lines. In: Journal of human genetics. 2013 ; Vol. 58, No. 1. pp. 16-20.
@article{41591fcfe05f44f78414f1ac9d5da5c9,
title = "Genotype instability during long-term subculture of lymphoblastoid cell lines",
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.",
author = "Oh, {Ji Hee} and Kim, {Young Jin} and Sanghoon Moon and Nam, {Hye Young} and Jeon, {Jae Pil} and Lee, {Jong Ho} and Lee, {Jong Young} and Cho, {Yoon Shin}",
year = "2013",
month = "1",
day = "1",
doi = "10.1038/jhg.2012.123",
language = "English",
volume = "58",
pages = "16--20",
journal = "Journal of Human Genetics",
issn = "1434-5161",
publisher = "Nature Publishing Group",
number = "1",

}

Oh, JH, Kim, YJ, Moon, S, Nam, HY, Jeon, JP, Lee, JH, Lee, JY & Cho, YS 2013, 'Genotype instability during long-term subculture of lymphoblastoid cell lines', Journal of human genetics, vol. 58, no. 1, pp. 16-20. https://doi.org/10.1038/jhg.2012.123

Genotype instability during long-term subculture of lymphoblastoid cell lines. / Oh, Ji Hee; Kim, Young Jin; Moon, Sanghoon; Nam, Hye Young; Jeon, Jae Pil; Lee, Jong Ho; Lee, Jong Young; Cho, Yoon Shin.

In: Journal of human genetics, Vol. 58, No. 1, 01.01.2013, p. 16-20.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Genotype instability during long-term subculture of lymphoblastoid cell lines

AU - Oh, Ji Hee

AU - Kim, Young Jin

AU - Moon, Sanghoon

AU - Nam, Hye Young

AU - Jeon, Jae Pil

AU - Lee, Jong Ho

AU - Lee, Jong Young

AU - Cho, Yoon Shin

PY - 2013/1/1

Y1 - 2013/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84873020917&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84873020917&partnerID=8YFLogxK

U2 - 10.1038/jhg.2012.123

DO - 10.1038/jhg.2012.123

M3 - Article

VL - 58

SP - 16

EP - 20

JO - Journal of Human Genetics

JF - Journal of Human Genetics

SN - 1434-5161

IS - 1

ER -