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Characterization of adenosine deaminase (ADA)‐negative B‐lymphoblastoid cells cocultured with ADA‐positive fibroblasts
Author(s) -
Fujita Masahiro,
Ito Kazuhiko,
Kawamoto Hiroshi,
Kashii Saburo,
Norioka Mihoko,
Monden Sumie,
Okuma Minoru
Publication year - 1993
Publication title -
european journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.904
H-Index - 84
eISSN - 1600-0609
pISSN - 0902-4441
DOI - 10.1111/j.1600-0609.1993.tb01921.x
Subject(s) - adenosine deaminase , lymphoblast , deoxyadenosine , microbiology and biotechnology , cell culture , fibroblast , chemistry , adenosine , biology , biochemistry , genetics
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 × 10 5 and 1 × 10 6 cells/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.