Assessing the impact of minimizing arginine conversion in fully defined SILAC culture medium in human embryonic stem cells.

ABSTRACT: We present a fully defined culture system (adapted Essential8^TM [E8^TM ] medium in combination with vitronectin) for human embryonic stem cells that can be used for SILAC purposes. Although a complete incorporation of the labels was observed after 4 days in culture, over 90% of precursors showed at least 10% conversion. To reduce this arginine conversion, E8^TM medium was modified by adding (1) l-proline, (2) l-ornithine, (3) N^ω -hydroxy-nor-l-arginine acetate, or by (4) lowering the arginine concentration. Reduction of arginine conversion was best obtained by adding 5 mM l-ornithine, followed by 3.5 mM l-proline and by lowering the arginine concentration in the medium to 99.5 μM. No major changes in pluripotency and cell amount could be observed f

SUBMITTER: Scheerlinck E 

PROVIDER: S-EPMC5096064 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature