title: Fast quantitative real-time PCR-based screening for common chromosomal aneuploidies in mouse embryonic stem cells.
abstract: Chromosomal integrity has been known for many years to affect the ability of mouse embryonic stem cells (mESCs) to contribute to the germline of chimeric mice. Abnormal chromosomes are generally detected by standard cytogenetic karyotyping. However, this method is expensive, time consuming, and often omitted prior to blastocyst injection, consequently reducing the frequency of mESC-derived offspring. Here, we show a fast, accurate, and inexpensive screen for identifying the two most common aneuploidies (Trisomy 8 and loss of chromosome Y) in genetically manipulated mESCs using quantitative real-time PCR (qPCR). Screening against these two aneuploidies significantly increases the fraction of normal mESC clones. Our method is extremely sensitive and can detect as low as 10% aneuploidy among a large population of mESCs. It greatly expedites the generation of mutant mice and provides a quick tool for assessing the aneuploidy percentages of any mESC line.
authors: Department of Biological Sciences, Hunter College and The Graduate Center Biochemistry, Biology and Biopsychology and Behavioral Neuroscience Programs, The City University of New York, New York, NY 10065, USA.
authors: Charlotte
authors: C
authors: D'Hulst
authors: Irena
authors: I
authors: Parvanova
authors: Delia
authors: D
authors: Tomoiaga
authors: Maria L
authors: ML
authors: Sapar
authors: Paul
authors: P
authors: Feinstein
mesh_terms: D000782:Aneuploidy; D000818:Animals; D002872:Chromosome Deletion; D002898:Chromosomes, Human, Pair 8; D003198:Computer Simulation; D053595:Embryonic Stem Cells; D007621:Karyotyping / Q000379:methods*; D051379:Mice; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D014314:Trisomy; D014998:Y Chromosome