Optimal phenotypic expression times for HPRT mutants induced in foreskin‐, skin‐, and lung‐derived human diploid fibroblasts

1 January 1983

journal article
research article
Published by Wiley in Environmental Mutagenesis

Vol. 5 (5) , 717-731
https://doi.org/10.1002/em.2860050510

Abstract

A series of paired lung- and skin-derived fibroblast cultures has been established from human embryonic tissues under carefully controlled, identical conditions, providing the unique opportunity to study differences between normal diploid fibroblast populations from skin and lung without the confusion of genetic differences between donors. To reliably assess differences in the induced mutation frequencies observed among different cell populations, optimal phenotypic expression times in the HPRT mutagenesis system were determined for neonatal foreskin, fetal skin, and fetal lung cultures. Cell populations were mutagenized with several doses of N-methyl-N'-nitro-N-nitrosoguanidine and were replated in 6-thioguanine selective medium at intervals over 14 days. Survivals following MNNG exposure ranged from 1.6% to 45.5%. For all doses and survivals tested a 7-day expression period was the optimal value for cultures from the three different tissue sources in six independent experiments. Mutant frequency data derived from untreated control populations confirmed that spontaneous mutations during the expression period contributed negligibly to the final mutant frequency. Differences between the mutation frequencies obtained using an in situ and a replating protocol were approximately twofold for lung-, skin-, and foreskin-derived cultures.

Keywords

This publication has 23 references indexed in Scilit:

The quantitation of TK^−/− and HGPRT⁻ mutants of L5178Y/TK^+/− mouse lymphoma cells at varying times post‐treatment
Environmental Mutagenesis, 1982
Conditions necessary for quantifying ethyl methanesulfonate-induced mutations to purine-analogue resistance in Chinese hamster V79 cells
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1978
Chemical mutagenesis of mammalian cells can be quantified
Nature, 1977
Dilution of hypoxanthine-guanine phosphoribosyl transferase and other factors affecting the frequency of 6-thioguanine resistance in Chinese hamster lung cells
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1977
X-ray-induced mutation to 6-thioguanine resistance in cultured human diploid fibroblasts
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1976
Linear dose —response relationships after prolonged expression times in V-79 Chinese hamster cells
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1976
Ethyl methanesulphonate mutagenesis with L5178Y mouse lymphoma cells: A comparison of ouabain, thioguanine and excess thymidine resistance
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1976
The use of correction factors in the determination of mutant frequencies in populations of human diploid skin fibroblasts
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1976
A mutational assay system for L5178Y mouse lymphoma cells, using hypoxanthine-guanine-phosphoribosyl-transferase (HGPRT) -deficiency as marker. The occurrence of a long expression time for mutations induced by X-rays and EMS
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, 1975
Detection of mycoplasma in cell cultures
Tissue Culture Association Manual, 1975