Characterization of adenosine deaminase (ADA)‐negative B‐lymphoblastoid cells cocultured with ADA‐positive fibroblasts

Masahiro Fujita, Kazuhiko Ito, Hiroshi Kawamoto, Saburo Kashii, Mihoko Norioka, Sumie Monden, Minoru Okuma

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Abstract: A cell line, BAD05, derived from B lymphocytes of an adenosine deaminase (ADA; EC 3,5,4,4)‐deficient patient could not proliferate in a serum‐free medium containing 100 μmol/l deoxyadenosine. When BAD05 was cultured with ADA‐positive fibroblasts, the proliferation of BAD05 was improved. BAD05 cell density increased when the initially mixed ratio of fibroblasts/BAD05 was 1/10 or higher, but decreased when the ratio was 1/20 or lower. Deoxyadenosine concentrations in the medium and ATP and deoxyATP (dATP) levels in the BAD05 were measured after 4 hours of coculture at initial BAD05 cell densities of 1 × 105and 1 × 106cells/ml. Deoxyadenosine concentrations in the medium decreased as the density of fibroblasts increased. The dATP level decreased as the mixed ratio rose. The ratio of fibroblasts/BAD05 rather than the cell density of fibroblasts had a larger effect on the dATP levels in BAD05. Under our experimental conditions, ADA‐negative cells proliferated well when the ratio of ADA‐positive cells/ADA‐negative cells was over 1/10.

Original languageEnglish
Pages (from-to)200-205
Number of pages6
JournalEuropean Journal of Haematology
Volume50
Issue number4
DOIs
Publication statusPublished - 04-1993

All Science Journal Classification (ASJC) codes

  • Hematology

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