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

(2016) PROTEOMICS. 16(20). p.2605-2614
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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-omega-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 mu M. No major changes in pluripotency and cell amount could be observed for the adapted E8 (TM) media with ornithine and proline. However, our subsequent ion mobility assisted data-independent acquisition (high-definition MS) proteome analysis cautions for ongoing changes in the proteome when aiming at longer term suppression of arginine conversion.
Keywords
AMINO-ACIDS, MASS-SPECTROMETRY, PROLIFERATION, PROTEOMICS, Arginine conversion, Cell culture, hESC, SILAC, Technology

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Chicago
Scheerlinck, Ellen, Katleen Van Steendam, Simon Daled, Elisabeth Govaert, Liesbeth Vossaert, Paulien Meert, Filip Van Nieuwerburgh, et al. 2016. “Assessing the Impact of Minimizing Arginine Conversion in Fully Defined SILAC Culture Medium in Human Embryonic Stem Cells.” Proteomics 16 (20): 2605–2614.
APA
Scheerlinck, Ellen, Van Steendam, K., Daled, S., Govaert, E., Vossaert, L., Meert, P., Van Nieuwerburgh, F., et al. (2016). Assessing the impact of minimizing arginine conversion in fully defined SILAC culture medium in human embryonic stem cells. PROTEOMICS, 16(20), 2605–2614.
Vancouver
1.
Scheerlinck E, Van Steendam K, Daled S, Govaert E, Vossaert L, Meert P, et al. Assessing the impact of minimizing arginine conversion in fully defined SILAC culture medium in human embryonic stem cells. PROTEOMICS. 2016;16(20):2605–14.
MLA
Scheerlinck, Ellen, Katleen Van Steendam, Simon Daled, et al. “Assessing the Impact of Minimizing Arginine Conversion in Fully Defined SILAC Culture Medium in Human Embryonic Stem Cells.” PROTEOMICS 16.20 (2016): 2605–2614. Print.
@article{8500529,
  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-omega-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 mu M. No major changes in pluripotency and cell amount could be observed for the adapted E8 (TM) media with ornithine and proline. However, our subsequent ion mobility assisted data-independent acquisition (high-definition MS) proteome analysis cautions for ongoing changes in the proteome when aiming at longer term suppression of arginine conversion.},
  author       = {Scheerlinck, Ellen and Van Steendam, Katleen and Daled, Simon and Govaert, Elisabeth and Vossaert, Liesbeth and Meert, Paulien and Van Nieuwerburgh, Filip and Van Soom, Ann and Peelman, Luc and De Sutter, Petra and Heindryckx, Bj{\"o}rn and Dhaenens, Maarten and Deforce, Dieter},
  issn         = {1615-9853},
  journal      = {PROTEOMICS},
  keyword      = {AMINO-ACIDS,MASS-SPECTROMETRY,PROLIFERATION,PROTEOMICS,Arginine conversion,Cell culture,hESC,SILAC,Technology},
  language     = {eng},
  number       = {20},
  pages        = {2605--2614},
  title        = {Assessing the impact of minimizing arginine conversion in fully defined SILAC culture medium in human embryonic stem cells},
  url          = {http://dx.doi.org/10.1002/pmic.201600174},
  volume       = {16},
  year         = {2016},
}

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