Selection of morphologically normal cell lines from polyoma-transformed BHK21/13 hamster fibroblasts

Abstract

A selective method was devised for the isolation of "revertants" from polyoma-transformed sublines derived from BHK21/13 Syrian hamster fibroblasts. A hybrid, polyploid subline was obtained by growing together, in mixed culture in the presence of aminopterin, two variant BHK21/13 sublines lacking either inosinic acid pyrophosphorylase or thymidine kinase. Whereas these variant sublines were resistant to 6-thioguanine or to 5-bromodeoxyuridine, the hybrid had regained sensitivity to both analogues. By plating a polyoma-transformed subline derived from this hybrid in the presence of 6-thioguanine, resistant clones were obtained with a frequency of about 10(-4). All of these surviving clones had a reduced chromosome complement and some of them had regained a normal phenotype.