DNA content of microcells prepared from rat kangaroo and mouse cells

Abstract

Enucleation of animal cells in which nuclear fragmentation (micronucleation) has been induced by treatment with mitotic inhibitors results in the formation of subdiploid microcells consisting of one or several micronuclei, some cytoplasm and surrounded by a plasma membrane. Microcells were prepared from rat kangaroo cells (12 chromosomes) and a polyoma virus transformed mouse cell line (61 chromosomes) and analysed for DNA content. Microspectrophotometric DNA measurements and the appearance of micronuclei at mitosis show that small micronuclei contain genetic information equivalent to single chromosomes. A large proportion of the micronuclei and the microcells, however, contains DNA corresponding to several chromosomes. Heterogeneous mixtures of microcells can be fractionated by a unit gravity sedimentation procedure so as to isolate the small microcells. These can afterwards be fused with intact normal or mutant cells.