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)