Characterization of adenosine deaminase (ADA)-negative B-lymphoblastoid cells cocultured with ADA-positive fibroblasts.
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 mumol/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 x 10(5) and 1 x 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.
Department of Internal Medicine, Faculty of Medicine, Kyoto University, Japan.
M
M
Fujita
K
K
Ito
H
H
Kawamoto
S
S
Kashii
M
M
Norioka
S
S
Monden
M
M
Okuma
D000243:Adenosine Deaminase / Q000172:deficiency* / Q000502:physiology; D000255:Adenosine Triphosphate / Q000378:metabolism; D001402:B-Lymphocytes / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D003839:Deoxyadenosines / Q000378:metabolism; D005347:Fibroblasts / Q000201:enzymology*; D006801:Humans; D007958:Leukocyte Count
D003838:Deoxyadenine Nucleotides; D003839:Deoxyadenosines; D000255:Adenosine Triphosphate; D000243:Adenosine Deaminase; C026600:2'-deoxyadenosine triphosphate