Capacity of robot handling for Epstein‐Barr virus transformation

Chang, I–Shou; Ko, Hui-Wen; Hwang, Shiaw-Min

doi:10.1111/j.1365-2184.2008.00568.x

Cited by 1 publication

(1 citation statement)

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“…To the cells suspended in above-described standard B cell culture medium, a 50% volume of B95-8 supernatant was added to become 1.0 × 10 6 cells/ml as the final concentration. For establishing and maintaining LCLs, 1% (v/v) phytohemagglutinin (Life Technologies) was added 25 . Cells were then cultured at 37 °C in 5% CO 2 , and one-third of the culture supernatant was replaced with fresh medium every 3-4 days.…”

Section: Methodsmentioning

confidence: 99%

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

Omi

Tokuda

Ikeda

et al. 2017

Sci Rep

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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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Section: Methodsmentioning

confidence: 99%