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

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  • Additional Information
    • Source:
      Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE
    • Publication Information:
      Original Publication: London : Nature Publishing Group, copyright 2011-
    • Subject Terms:
      Polymorphism, Single Nucleotide*; Genome, Human/*genetics ; Leukocytes/*metabolism ; Leukocytes, Mononuclear/*metabolism ; Lymphocytes/*metabolism; Cell Line, Transformed ; Genotype ; Herpesvirus 4, Human/physiology ; Humans ; Leukocytes/cytology ; Leukocytes/virology ; Leukocytes, Mononuclear/cytology ; Leukocytes, Mononuclear/virology ; Lymphocytes/virology ; Oligonucleotide Array Sequence Analysis ; Reproducibility of Results
    • 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.
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    • Publication Date:
      Date Created: 20170309 Date Completed: 20181106 Latest Revision: 20181113
    • Publication Date:
      20250114
    • Accession Number:
      PMC5341036
    • Accession Number:
      10.1038/srep43833
    • Accession Number:
      28272413