title: An enhanced chemically defined SILAC culture system for quantitative proteomics study of human embryonic stem cells.
abstract: Stable isotope labeling by SILAC-based quantitative proteomics analysis provides an unprecedented tool for the study of mechanisms underlying the self-renewal and differentiation of human embryonic stem cells (hESCs). While we recently reported a chemically defined SILAC culture system specific for a rare cell proteomic reactor (R. Tian et al., Mol. Cell. Proteomics 2011, 10, M110.000679), total hESC yield, prolonged self-renewal capacity (i.e.<12 days), and laborious procedure remain substantial hurdles for its conventional application in hESC studies. Here, we devised an enhanced SILAC culture system consisting of a new chemically defined SILAC-medium and a novel culture protocol. As a result, with much less culture maneuvers, approximately 40-fold greater hESCs were produced than the system reported previously. Moreover, the enhanced SILAC culture system was sufficient to support the self-renewal of hESCs for >60 days and was also highly reproducible. As such, it provides a new platform that can be readily adapted by general laboratory for further comprehensive SILAC-based proteomics analysis of hESCs and induced pluripotent stem cells.
authors.affiliation: Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, Canada.
authors.forename: Shuai
authors.initials: S
authors.lastname: Wang
authors.forename: Ruijun
authors.initials: R
authors.lastname: Tian
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.forename: Daniel
authors.initials: D
authors.lastname: Figeys
authors.forename: Lisheng
authors.initials: L
authors.lastname: Wang
mesh_terms: D018929:Cell Culture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D007553:Isotope Labeling; D040901:Proteomics / Q000379:methods*