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An enhanced chemically defined SILAC culture system for quantitative proteomics study of human embryonic stem cells
Author(s) -
Wang Shuai,
Tian Ruijun,
Li Li,
Figeys Daniel,
Wang Lisheng
Publication year - 2011
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201100052
Subject(s) - stable isotope labeling by amino acids in cell culture , embryonic stem cell , proteomics , quantitative proteomics , stem cell , chemistry , microbiology and biotechnology , computational biology , biology , biochemistry , gene
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.