Protocols for cytogenetic studies of human embryonic stem cells.
All cultured cells develop chromosome changes over time, including cultures of human embryonic stem cells (hESC), but only those cells with adaptive chromosomes changes survive. The most frequent chromosome changes in hESC cultures are trisomy 12 and trisomy 17. Cells with these trisomies are indistinguishable from normal cells by appearance and also demonstrate typical markers of pluripotency, making them difficult to identify without cytogenetic analysis. Early detection of these cells is essential since cells with trisomy 12 and 17 can replace the normal cell population in 5-10 passages. Cytogenetic analysis using G-banding is considered to be the gold standard for detecting chromosome abnormalities and, when used in combination with interphase FISH, provides a sensitive method for early detection of cytogenetic aberrations, such as full and partial trisomies of chromosomes 12 and 17. The following discussion describes the cytogenetic methods used in our laboratory to study cultured hESCs, along with recommendations for integrating these methods into a plan for routine cell line quality control.
Meisner
Lorraine Faxon
LF
Department of Population Health Sciences, University of Wisconsin, and Cell Line Genetics, LLC, 510 Charmany Drive, Suite 254, Madison, WI 53719, USA. lmeisner@clgenetics.com
Johnson
Julie A
JA
D002871:Chromosome Banding; D002881:Chromosomes, Human, Pair 12 / Q000648:ultrastructure; D002886:Chromosomes, Human, Pair 17 / Q000648:ultrastructure; D003582:Cytogenetics / Q000379:methods*; D033041:Embryo Research; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D014314:Trisomy / Q000175:diagnosis*