Cell line/type | ESCs (Embryonic Stem Cells) |
---|---|
Species | Human |
Animal free | Unspecified |
Product | mTeSR1 Liberski et al. 2013 |
Liberski, A. R., Al-Noubi, M. N., Rahman, Z. H., Halabi, N. M., Dib, S. S., Al-Mismar, R., ... & Rafii, A. (2013). Adaptation of a commonly used, chemically defined medium for human embryonic stem cells to stable isotope labeling with amino acids in cell culture. Journal of proteome research, 12(7), 3233-3245. In this study, a feeder-free SILAC culture system based on a customized version of a commonly used, chemically defined hESC medium developed by Ludwig et al. and commercially available as mTeSR1 is demonstrated. A system for metabolic (SILAC) labeling of human embryonic stem cells was established in a modified version of the widely published and commonly used mTeSR1 medium. To titrate the amount of SILAC amino acids required by hESCs, 1.66 × 105 cells per well of a six-well plate were plated into standard mTeSR1 and customized mTeSR1 devoid of arginine and lysine supplemented with 100% (0.548 mM Arg, 0.391 mM Lys), 50%, 33%, 25% and 0% of the arginine and lysine concentrations found in the standard medium. Using 25% of the arginine and lysine concentrations provided by the standard medium, essentially complete labeling was observed after 96 h of culture without any obvious reduction in proliferative capacity as compared to the default medium. This medium, together with adjustments to the culturing protocol, facilitates reproducible labeling that is easily scalable to the protein amounts required by proteomic workflows. It greatly enhances the usability of quantitative proteomics as a tool for the study of mechanisms underlying hESCs differentiation and self-renewal. mTeSR1 is a commercial product. The precise formulation is proprietary. https://pubs.acs.org/doi/pdfplus/10.1021/pr400099j |
|
Source | Literature - modified commercial product |
Chemically defined > Yes | Yes |
Antibiotics free > No | No |