TY - JOUR UR - http://lib.ugent.be/catalog/pug01:8500529 ID - pug01:8500529 LA - eng TI - Assessing the impact of minimizing arginine conversion in fully defined SILAC culture medium in human embryonic stem cells PY - 2016 JO - (2016) PROTEOMICS SN - 1615-9853 SN - 1615-9861 PB - 2016 AU - Scheerlinck, Ellen UGent 002005460549 802000955989 AU - Van Steendam, Katleen UGent 002001216696 802000085417 AU - Daled, Simon FW01 000100705497 AU - Govaert, Elisabeth UGent 000080607404 802001653480 AU - Vossaert, Liesbeth UGent 002004110229 802000632455 AU - Meert, Paulien UGent 000060628333 802001079766 AU - Van Nieuwerburgh, Filip FW01 001994454483 AU - Van Soom, Ann DI08 801001060387 0000-0001-5010-6311 AU - Peelman, Luc DI07 801000721291 0000-0003-3853-5310 AU - De Sutter, Petra GE38 UZGent 001980047054 801000688454 0000-0002-8792-3814 AU - Heindryckx, Björn GE38 UZGent 001994280792 AU - Dhaenens, Maarten FW01 001998244456 AU - Deforce, Dieter FW01 801000962680 0000-0002-0635-661X AB - 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. ER -Download RIS file
| 00000nam^a2200301^i^4500 | |||
| 001 | 8500529 | ||
| 005 | 20180816075816.0 | ||
| 008 | 170105s2016------------------------eng-- | ||
| 022 | a 1615-9853 | ||
| 022 | a 1615-9861 | ||
| 024 | a 000387167700002 2 wos | ||
| 024 | a 1854/LU-8500529 2 handle | ||
| 024 | a 10.1002/pmic.201600174 2 doi | ||
| 040 | a UGent | ||
| 245 | a Assessing the impact of minimizing arginine conversion in fully defined SILAC culture medium in human embryonic stem cells | ||
| 260 | c 2016 | ||
| 520 | a 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. | ||
| 598 | a A1 | ||
| 100 | a Scheerlinck, Ellen u UGent 0 002005460549 0 802000955989 0 974229414915 9 0510100E-F0EE-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a Van Steendam, Katleen u UGent 0 002001216696 0 802000085417 0 971069125310 9 F88ED0CC-F0ED-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a Daled, Simon u FW01 0 000100705497 0 802002416346 9 2EC22C3E-F0EE-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a Govaert, Elisabeth u UGent 0 000080607404 0 802001653480 0 975095963203 9 149DEA28-F0EE-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a Vossaert, Liesbeth u UGent 0 002004110229 0 802000632455 0 976131554401 9 037A9F70-F0EE-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a Meert, Paulien u UGent 0 000060628333 0 802001079766 0 976191516565 9 029F8890-F0EE-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a Van Nieuwerburgh, Filip u FW01 0 001994454483 0 801001335829 9 F5BC58D8-F0ED-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a Van Soom, Ann u DI08 0 801001060387 0 0000-0001-5010-6311 9 F4F831BA-F0ED-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a Peelman, Luc u DI07 0 801000721291 0 0000-0003-3853-5310 9 F469A738-F0ED-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a De Sutter, Petra u GE38 u UZGent 0 001980047054 0 801000688454 0 0000-0002-8792-3814 9 F45AB642-F0ED-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a Heindryckx, Björn u GE38 u UZGent 0 001994280792 0 801001342701 9 F5C0698C-F0ED-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a Dhaenens, Maarten u FW01 0 001998244456 0 801002046454 9 F7EE877A-F0ED-11E1-A9DE-61C894A0A6B4 | ||
| 700 | a Deforce, Dieter u FW01 0 801000962680 0 0000-0002-0635-661X 9 F4D02620-F0ED-11E1-A9DE-61C894A0A6B4 | ||
| 650 | a Biology and Life Sciences | ||
| 653 | a AMINO-ACIDS | ||
| 653 | a MASS-SPECTROMETRY | ||
| 653 | a PROLIFERATION | ||
| 653 | a PROTEOMICS | ||
| 653 | a Arginine conversion | ||
| 653 | a Cell culture | ||
| 653 | a hESC | ||
| 653 | a SILAC | ||
| 653 | a Technology | ||
| 773 | t PROTEOMICS g Proteomics. 2016. 16 (20) p.2605-2614 q 16:20<2605 | ||
| 856 | 3 Full Text u https://biblio.ugent.be/publication/8500529/file/8500535 z [ugent] y Scheerlinck_et_al-2016-PROTEOMICS.pdf | ||
| 920 | a article | ||
| Z30 | x GE 1 GE04 | ||
| 922 | a UGENT-GE | ||
| Z30 | x FW 1 FW01 | ||
| 922 | a UGENT-FW | ||
| Z30 | x DI 1 DI07 | ||
| 922 | a UGENT-DI | ||
| Z30 | x DI 1 DI08 | ||
| 922 | a UGENT-DI | ||
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