Establishment of lung fibroblastic cell lines from a non-human primate Tupaia belangeri and their use in a forward gene mutation assay at the hypoxanthine-guanine phosphoribosyl transferase locus.
The cells obtained from a lung of a new-born male Tupaia belangeri were maintained in mass culture for greater than 400 days. After 55 population doubling levels (100 days in culture), three cell lines were separately established; these lines showed constant growth properties. One line, designated as T-23, was used for a mutation assay. The T-23 cells showed an absolute plating efficiency of 30-50%, and a population doubling time of 18-19 h in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum. The cells had a modal chromosome number of 62 (pseudodiploid) with the loss of a chromosome and the gain of an unidentified one. T-23 cells, like human cells, were much more susceptible to ouabain than mouse cells but relatively less susceptible to 8-azaguanine, while, unlike human cells, they were less sensitive to 6-thioguanine (6TG). N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG) was less, but 4-nitroquinoline-1-oxide (4NQO) was more toxic to T-23 cells than to human or mouse cells. Benzo[a]pyrene-induced toxicity was almost comparable among the cell types. For the mutation assay, we chose 6TG-resistance (100 microM) as a marker. The optimal expression time (8-13 days) and cell density at selection to eliminate metabolic cooperation (2 x 10(4) cells/60-mm dish) were determined. Some of the cells selected with 6TG showed less than 0.4% of the total incorporation of [14C]hypoxanthine into wild-type cells, suggesting the mutants under selection were affected at the hypoxanthine-guanine phosphoribosyl transferase locus.(ABSTRACT TRUNCATED AT 250 WORDS)
Taketomi
M
M
Section of Cell Biology and Cytogenetics, Japan Tobacco Inc., Kanagawa.
Nishi
Y
Y
Ohkawa
Y
Y
Inui
N
N
D000818:Animals; D000831:Animals, Newborn; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004351:Drug Resistance / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000187:drug effects; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008168:Lung / Q000166:cytology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000494:pharmacology; D013866:Thioguanine / Q000494:pharmacology*; D014417:Tupaia / Q000235:genetics*; D014418:Tupaiidae / Q000235:genetics*
D009153:Mutagens; D007041:Hypoxanthine Phosphoribosyltransferase; D013866:Thioguanine