Application of spectral karyotyping to the analysis of the human chromosome complement of interspecies somatic cell hybrids.
Mouse-human somatic cell hybrids have been extensively used in the molecular genetic dissection of human disease-related chromosome rearrangements because of their ability to selectively and randomly eliminate human chromosomes. This technology allows the isolation of structural chromosome abnormalities, which then allows determination of the precise molecular address of chromosome breakpoints associated with deletions and translocations, down to the nucleotides involved. The main confounding problem with the analysis of somatic cell hybrids is determining the exact chromosome complement unequivocally and quickly. Spectral karyotyping can identify each of the individual human chromosomes in a normal metaphase spread, as well as structural chromosome rearrangements-although, because of potential cross-hybridization between the human probe and mouse DNA sequences during the hybridization reaction, it has not been determined whether the same analysis will selectively identify human chromosomes on a mouse background. We show (to our knowledge, for the first time) that, under modified conditions of chromosomal in situ suppression hybridization, the standard spectral karyotyping probe does not cross-react with mouse chromosomes and can be used to identify subtle structurally rearranged chromosomes in hybrid cells. This analysis allows for the rapid and unequivocal identification of the human chromosome complement in these hybrids, as well as structural chromosome rearrangements that occur between mouse and human chromosomes that might otherwise confound the analysis.
Matsui
Sei-ichi
S
Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, NY 14263, USA.
Faitar
Silviu L
SL
Rossi
Michael R
MR
Cowell
John K
JK
D016475:3T3 Cells; D000818:Animals; D002459:Cell Fusion; D002877:Chromosomes, Human; D015342:DNA Probes; D005910:Glioma / Q000235:genetics; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology / Q000502:physiology*; D008677:Metaphase / Q000235:genetics; D051379:Mice; D032681:Spectral Karyotyping / Q000379:methods*; D014178:Translocation, Genetic; D014407:Tumor Cells, Cultured
D015342:DNA Probes