title: Multicolor karyotype analyses of mouse embryonic stem cells.
abstract: The manipulation of embryonic stem (ES) cells to introduce directional genetic changes into the genome of mice has become an important tool in biomedical research. Monitoring of cell morphology before and after DNA manipulation and special culture conditions are a prerequisite to preserve the pluripotent properties of ES cells and thus their ability to generate chimera and effective germline transmission (GLT). It has been reported that prolonged cell culturing may affect the diploid chromosomal composition of cells and therefore the percentage of chimerism and GLT. Herein, we report multicolor-fluorescence in situ hybridization (M-FISH) analysis of four different ES cell lines/clones. Although the morphology of all four ES cell lines/clones appeared normal and all four expressed the early markers Oct-3/4 and Nanog, two cell lines presented consistent numerical and structural chromosome aberrations. We demonstrate that M-FISH is a sensitive and accurate method for a comprehensive karyotype analysis of ES cells and may minimize time, costs, and disappointments due to inadequate ES cell sources.
authors.affiliation: Institute of Human Genetics, University of Heidelberg, D-69120 Heidelberg, Germany.
authors.forename: Jianli
authors.initials: J
authors.lastname: Guo
authors.forename: Anna
authors.initials: A
authors.lastname: Jauch
authors.forename: Holtgreve-Grez
authors.initials: HG
authors.lastname: Heidi
authors.forename: Brigitte
authors.initials: B
authors.lastname: Schoell
authors.forename: Dorothee
authors.initials: D
authors.lastname: Erz
authors.forename: Martina
authors.initials: M
authors.lastname: Schrank
authors.forename: Johannes W G
authors.initials: JW
authors.lastname: Janssen
mesh_terms: D000818:Animals; D033481:Chromosomes, Mammalian / Q000235:genetics*; D017931:DNA Primers; D004622:Embryo, Mammalian / Q000166:cytology*; D005069:Evaluation Studies as Topic; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007621:Karyotyping / Q000379:methods*; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction; D039901:Totipotent Stem Cells / Q000166:cytology*
chemical_list: D017931:DNA Primers