Efficient and reliable establishment of lymphoblastoid cell lines by Epstein-Barr virus transformation from a limited amount of peripheral blood View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2017-03-08

AUTHORS

Natsue Omi, Yuichi Tokuda, Yoko Ikeda, Morio Ueno, Kazuhiko Mori, Chie Sotozono, Shigeru Kinoshita, Masakazu Nakano, Kei Tashiro

ABSTRACT

Lymphoblastoid cell lines (LCLs) transformed by Epstein-Barr virus (EBV) serve as an unlimited resource of human genomic DNA. The protocol that is widely used to establish LCLs involves peripheral blood mononuclear cell isolation by density gradient centrifugation, however, that method requires as much as 5 ml of peripheral blood. In this study, in order to provide a more simple and efficient method for the generation of LCLs, we developed a new protocol using hemolytic reaction to enrich white blood cells for EBV transformation and found that the hemolytic protocol successfully generated LCLs from a small volume (i.e., 0.1 ml) of peripheral blood. To assess the quality of genomic DNA extracted from LCLs established by the hemolytic protocol (LCL-hemolytic), we performed single nucleotide polymorphism (SNP) microarray genotyping using the GeneChip® 100 K Array Set (Affymetrix, Inc.). The concordances of the SNP genotyping resulting from genomic DNA from LCL-hemolytic (99.92%) were found to be as good as the technical replicate (99.90%), and Kappa statistics results confirmed the reliability. The findings of this study reveal that the hemolytic protocol is a simple and reliable method for the generation of LCLs, even from a small volume of peripheral blood. More... »

PAGES

43833

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/srep43833

DOI

http://dx.doi.org/10.1038/srep43833

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1084131738

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/28272413


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