Development of a robust method for establishing B cell lines using Epstein-Barr virus

Inaho Danjoh, Hiyori Sone, Ryoko Shirota, Takashi Hiroyama, Yukio Nakamura

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

B lymphoblastoid cell lines (B-LCLs) are generally established from B lymphocytes by infection with Epstein- Barr virus (EBV). As their genomic structure is stable in culture, B-LCLs are a valuable resource for many types of analysis. The efficiency of establishing B-LCLs from freshly obtained blood samples from healthy individuals is almost 100 %; however, for blood samples stored inappropriately after collection or held in long-term storage as peripheral blood mononuclear cells (PBMCs) in liquid nitrogen, the efficiency of B-LCL establishment can be considerably lower. To date, we have established more than 550 B-LCLs from685 PBMC samples that have been stored in liquid nitrogen for over 20 yr. The PBMCs were prepared from blood samples donated by individuals belonging to native minority ethnic groups in outlying regions of South America and elsewhere. The establishment of B-LCLs from this material is difficult, and failure results in the waste of valuable and rare samples. We sought to improve our success rate for establishing B-LCLs from these difficult and irreplaceable samples by a detailed examination of each step of the process. The analysis showed that two parameters were particularly critical to the success rate: the density of the PBMCs plated after EBV infection and the EBV titer. These observations shed light on cases where establishment of B-LCLs was hard due to the small number of PBMCs or damage to the cells.

Original languageEnglish
Pages (from-to)393-402
Number of pages10
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume48
Issue number7
DOIs
Publication statusPublished - 2012 Aug 1
Externally publishedYes

Keywords

  • B cell line
  • EBV
  • Ethnic group
  • Genome analysis
  • South America

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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