#METABOLOMICS WORKBENCH leilirohani_20200508_183528 DATATRACK_ID:2003 STUDY_ID:ST001377 ANALYSIS_ID:AN002297 PROJECT_ID:PR000942
VERSION             	1
CREATED_ON             	May 13, 2020, 9:40 am
#PROJECT
PR:PROJECT_TITLE                 	Inferring metabolism dynamics between naïve and primed human pluripotent stem
PR:PROJECT_TITLE                 	cells in the stirred suspension bioreactor
PR:PROJECT_TYPE                  	Targeted and untargeted MS
PR:PROJECT_SUMMARY               	We used high-resolution LC-MS targeted metabolomics to screen for extracellular
PR:PROJECT_SUMMARY               	metabolites in the media of bioreactor-cultured, naïve and primed human
PR:PROJECT_SUMMARY               	pluripotent stem cells.
PR:INSTITUTE                     	University of Calgary
PR:DEPARTMENT                    	Biochemistry and Molecular Biology
PR:LABORATORY                    	Stem Cell Research
PR:LAST_NAME                     	Rohani
PR:FIRST_NAME                    	Leili
PR:ADDRESS                       	405J, 1919 University Drive, NW
PR:EMAIL                         	leili.rohanisarvesta@ucalgary.ca
PR:PHONE                         	5879681647
#STUDY
ST:STUDY_TITLE                   	Stirred suspension bioreactors maintain naïve pluripotency of human pluripotent
ST:STUDY_TITLE                   	stem cells (hPSCs)
ST:STUDY_SUMMARY                 	Although cell therapies require large numbers of quality-controlled hPSCs,
ST:STUDY_SUMMARY                 	existing technologies are limited in their ability to efficiently grow and scale
ST:STUDY_SUMMARY                 	stem cells. We report here that cell-state conversion from primed-to-naïve
ST:STUDY_SUMMARY                 	pluripotency enhances the biomanufacturing of hPSCs. Naïve hPSCs exhibit
ST:STUDY_SUMMARY                 	superior growth kinetics and aggregate formation characteristics in stirred
ST:STUDY_SUMMARY                 	suspension bioreactors compared to their primed counterparts. Moreover, we
ST:STUDY_SUMMARY                 	demonstrate the role of the bioreactor mechanical environment in the maintenance
ST:STUDY_SUMMARY                 	of naïve pluripotency, through transcriptomic enrichment of mechano-sensing
ST:STUDY_SUMMARY                 	signaling for cells in the bioreactor along with a decrease in expression of
ST:STUDY_SUMMARY                 	lineage-specific and primed pluripotency hallmarks. Bioreactor-cultured, naïve
ST:STUDY_SUMMARY                 	hPSCs express epigenetic regulatory transcripts associated with naïve
ST:STUDY_SUMMARY                 	pluripotency, and display hallmarks of X-chromosome reactivation. They exhibit
ST:STUDY_SUMMARY                 	robust production of naïve pluripotency metabolites and display reduced
ST:STUDY_SUMMARY                 	expression of primed pluripotency cell surface markers. We also show that these
ST:STUDY_SUMMARY                 	cells retain the ability to undergo targeted differentiation into beating
ST:STUDY_SUMMARY                 	cardiomyocytes, hepatocytes, and neural rosettes. They additionally display
ST:STUDY_SUMMARY                 	faster kinetics of teratoma formation compared to their primed counterparts.
ST:STUDY_SUMMARY                 	Naïve bioreactor hPSCs also retain structurally stable chromosomes. Our
ST:STUDY_SUMMARY                 	research corroborates that converting hPSCs to the naïve state enhances hPSC
ST:STUDY_SUMMARY                 	manufacturing and indicates a potentially important role for the bioreactor’s
ST:STUDY_SUMMARY                 	mechanical environment in maintaining naïve pluripotency.
ST:INSTITUTE                     	University of Calgary
ST:DEPARTMENT                    	Biochemistry and Molecular Biology
ST:LABORATORY                    	Stem Cell Research
ST:LAST_NAME                     	Rohani
ST:FIRST_NAME                    	Leili
ST:ADDRESS                       	405J, 1919 University Drive, NW
ST:EMAIL                         	leili.rohanisarvesta@ucalgary.ca
ST:PHONE                         	+1 5879681647
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Blank_mid6	Cell line:none | Status:none	RAW_FILE_NAME=2019_10_07LR_RPIP15_Blank_mid6.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Blank_mid7	Cell line:none | Status:none	RAW_FILE_NAME=2019_10_07LR_RPIP15_Blank_mid7.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Blank_mid8	Cell line:none | Status:none	RAW_FILE_NAME=2019_10_07LR_RPIP15_Blank_mid8.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Blank_mid9	Cell line:none | Status:none	RAW_FILE_NAME=2019_10_07LR_RPIP15_Blank_mid9.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Blank_mid10	Cell line:none | Status:none	RAW_FILE_NAME=2019_10_07LR_RPIP15_Blank_mid10.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H1-M1_1	Cell line:H1 | Status:day 1	RAW_FILE_NAME=2019_10_07LR_RPIP15_H1_M1_1.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H1-M1_2	Cell line:H1 | Status:day 2	RAW_FILE_NAME=2019_10_07LR_RPIP15_H1_M1_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H1-M2_1	Cell line:H1 | Status:day 3	RAW_FILE_NAME=2019_10_07LR_RPIP15_H1_M2_1.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H1-M2_2	Cell line:H1 | Status:day 4	RAW_FILE_NAME=2019_10_07LR_RPIP15_H1_M2_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H1-M3_1	Cell line:H1 | Status:day 3	RAW_FILE_NAME=2019_10_07LR_RPIP15_H1_M3_1.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H1-M3_2	Cell line:H1 | Status:day 3	RAW_FILE_NAME=2019_10_07LR_RPIP15_H1_M3_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H1-M4_1	Cell line:H1 | Status:day 4	RAW_FILE_NAME=2019_10_07LR_RPIP15_H1_M4_1.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H1-M4_2	Cell line:H1 | Status:day 4	RAW_FILE_NAME=2019_10_07LR_RPIP15_H1_M4_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H1-M5_1	Cell line:H1 | Status:day 5	RAW_FILE_NAME=2019_10_07LR_RPIP15_H1_M5_1.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H1-M5_2	Cell line:H1 | Status:day 5	RAW_FILE_NAME=2019_10_07LR_RPIP15_H1_M5_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H9-naive-bioreactor-1-	Cell line:H9 | Status:naive	RAW_FILE_NAME=2019_10_07LR_RPIP15_H9_naive_bioreactor_1_.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H9-naive-bioreactor-2	Cell line:H9 | Status:naive	RAW_FILE_NAME=2019_10_07LR_RPIP15_H9_naive_bioreactor_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H9-naive-bioreactor-3	Cell line:H9 | Status:naive	RAW_FILE_NAME=2019_10_07LR_RPIP15_H9_naive_bioreactor_3.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H9-primed-bioreactor-1	Cell line:H9 | Status:primed	RAW_FILE_NAME=2019_10_07LR_RPIP15_H9_primed_bioreactor_1_.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H9-primed-bioreactor-2	Cell line:H9 | Status:primed	RAW_FILE_NAME=2019_10_07LR_RPIP15_H9_primed_bioreactor_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_H9-primed-bioreactor-3	Cell line:H9 | Status:primed	RAW_FILE_NAME=2019_10_07LR_RPIP15_H9_primed_bioreactor_3.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H1-M1_1	Cell line:H1 | Status:day 1	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H1_M1_1.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H1-M1_2	Cell line:H1 | Status:day 2	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H1_M1_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H1-M2_1	Cell line:H1 | Status:day 3	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H1_M2_1.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H1-M2_2	Cell line:H1 | Status:day 4	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H1_M2_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H1-M3_1	Cell line:H1 | Status:day 3	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H1_M3_1.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H1-M3_2	Cell line:H1 | Status:day 3	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H1_M3_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H1-M4_1	Cell line:H1 | Status:day 4	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H1_M4_1.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H1-M4_2	Cell line:H1 | Status:day 4	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H1_M4_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H1-M5_1	Cell line:H1 | Status:day 5	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H1_M5_1.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H1-M5_2	Cell line:H1 | Status:day 5	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H1_M5_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H9-naive-bioreactor-1	Cell line:H9 | Status:naive	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H9_naive_bioreactor_1_.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H9-naive-bioreactor-2	Cell line:H9 | Status:naive	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H9_naive_bioreactor_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H9-naive-bioreactor-3	Cell line:H9 | Status:naive	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H9_naive_bioreactor_3.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H9-primed-bioreactor-1	Cell line:H9 | Status:primed	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H9_primed_bioreactor_1_.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H9-primed-bioreactor-2	Cell line:H9 | Status:primed	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H9_primed_bioreactor_2.raw
SUBJECT_SAMPLE_FACTORS           	-	2019_09_13_RPIP15_Ref-Media-H9-primed-bioreactor-3	Cell line:H9 | Status:primed	RAW_FILE_NAME=2019_10_07LR_RPIP15_Ref_Media_H9_primed_bioreactor_3.raw
#COLLECTION
CO:COLLECTION_SUMMARY            	Extracellular metabolite dynamics was analyzed during primed to naïve
CO:COLLECTION_SUMMARY            	conversion in the bioreactor. For this, the media of H1 hESCs in the bioreactor
CO:COLLECTION_SUMMARY            	was gradually switched from mTESR1 to RSeT media within 6 days of culture (D2 to
CO:COLLECTION_SUMMARY            	D6) and the media was collected every day for the analysis. For H9 hESCs, the
CO:COLLECTION_SUMMARY            	extracellular metabolites were analyzed in the spent media of primed and
CO:COLLECTION_SUMMARY            	established naïve (P5) hPSCs in the bioreactor, and the media of each primed or
CO:COLLECTION_SUMMARY            	naïve hPSCs was collected at day four of bioreactor culture.
CO:SAMPLE_TYPE                   	Stem cells
#TREATMENT
TR:TREATMENT_SUMMARY             	For H1 hESCs, the extracellular metabolite dynamics was analyzed during primed
TR:TREATMENT_SUMMARY             	to naïve conversion in the bioreactor. For this, the media of H1 hESCs in the
TR:TREATMENT_SUMMARY             	bioreactor was gradually switched from mTESR1 to RSeT media within 6 days of
TR:TREATMENT_SUMMARY             	culture (D2 to D6) and the media was collected every day for the analysis. For
TR:TREATMENT_SUMMARY             	H9 hESCs, the extracellular metabolites were analyzed in the spent media of
TR:TREATMENT_SUMMARY             	primed and established naïve (P5) hPSCs in the bioreactor, and the media of
TR:TREATMENT_SUMMARY             	each primed or naïve hPSCs was collected at day four of bioreactor culture.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	The metabolite extraction was started using LC-MS or HPLC grade methanol
SP:SAMPLEPREP_SUMMARY            	(Sigma-Aldrich, 1.06035). Briefly, a 950 μL of pre-chilled 50% MeOH/H2O was
SP:SAMPLEPREP_SUMMARY            	added to a 50 μL of the bioreactor-collected media (making a D20 dilution), and
SP:SAMPLEPREP_SUMMARY            	the samples were incubated on ice for 30 min to allow for full extraction.
SP:SAMPLEPREP_SUMMARY            	Macromolecules were then pelleted by centrifugation at max speed (~18,000-21,000
SP:SAMPLEPREP_SUMMARY            	g) for 10 mins in a bench top centrifuge (preferably chilled) to extract the
SP:SAMPLEPREP_SUMMARY            	supernatant. Further, the extracted samples were stored in -80 prior to running
SP:SAMPLEPREP_SUMMARY            	HPLC-MS.
SP:PROCESSING_STORAGE_CONDITIONS 	Room temperature
SP:EXTRACTION_METHOD             	50% Methanol in water
SP:EXTRACT_STORAGE               	On ice
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Agilent Zorbax RRHT SB-C18 (50 x 2.1 mm, 1.8 um)
CH:FLOW_RATE                     	0.6 ml/min
CH:SOLVENT_A                     	10mM tributylamine, 10mM acetate pH 7.5 in 97/3% (v/v) MS grade water/methanol
CH:SOLVENT_B                     	MS grade acetonitrile
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:LABORATORY_NAME               	Ian Lewis
AN:OPERATOR_NAME                 	Ryan Groves
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive HF hybrid Orbitrap
MS:INSTRUMENT_TYPE               	Single quadrupole
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	The mass spectrometer was run in negative full scan mode at a resolution of
MS:MS_COMMENTS                   	140,000 scanning from 50-750m/z. Data was processed using MAVEN.
MS:CAPILLARY_TEMPERATURE         	275
MS:SPRAY_VOLTAGE                 	2.50 kV
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	relative signal intensity compared to media only
MS_METABOLITE_DATA_START
Samples	2019_09_13_RPIP15_Blank_mid6	2019_09_13_RPIP15_Blank_mid7	2019_09_13_RPIP15_Blank_mid8	2019_09_13_RPIP15_Blank_mid9	2019_09_13_RPIP15_Blank_mid10	2019_09_13_RPIP15_H1-M1_1	2019_09_13_RPIP15_H1-M1_2	2019_09_13_RPIP15_H1-M2_1	2019_09_13_RPIP15_H1-M2_2	2019_09_13_RPIP15_H1-M3_1	2019_09_13_RPIP15_H1-M3_2	2019_09_13_RPIP15_H1-M4_1	2019_09_13_RPIP15_H1-M4_2	2019_09_13_RPIP15_H1-M5_1	2019_09_13_RPIP15_H1-M5_2	2019_09_13_RPIP15_H9-naive-bioreactor-1-	2019_09_13_RPIP15_H9-naive-bioreactor-2	2019_09_13_RPIP15_H9-naive-bioreactor-3	2019_09_13_RPIP15_H9-primed-bioreactor-1	2019_09_13_RPIP15_H9-primed-bioreactor-2	2019_09_13_RPIP15_H9-primed-bioreactor-3	2019_09_13_RPIP15_Ref-Media-H1-M1_1	2019_09_13_RPIP15_Ref-Media-H1-M1_2	2019_09_13_RPIP15_Ref-Media-H1-M2_1	2019_09_13_RPIP15_Ref-Media-H1-M2_2	2019_09_13_RPIP15_Ref-Media-H1-M3_1	2019_09_13_RPIP15_Ref-Media-H1-M3_2	2019_09_13_RPIP15_Ref-Media-H1-M4_1	2019_09_13_RPIP15_Ref-Media-H1-M4_2	2019_09_13_RPIP15_Ref-Media-H1-M5_1	2019_09_13_RPIP15_Ref-Media-H1-M5_2	2019_09_13_RPIP15_Ref-Media-H9-naive-bioreactor-1	2019_09_13_RPIP15_Ref-Media-H9-naive-bioreactor-2	2019_09_13_RPIP15_Ref-Media-H9-naive-bioreactor-3	2019_09_13_RPIP15_Ref-Media-H9-primed-bioreactor-1	2019_09_13_RPIP15_Ref-Media-H9-primed-bioreactor-2	2019_09_13_RPIP15_Ref-Media-H9-primed-bioreactor-3
Factors	Cell line:none | Status:none	Cell line:none | Status:none	Cell line:none | Status:none	Cell line:none | Status:none	Cell line:none | Status:none	Cell line:H1 | Status:day 1	Cell line:H1 | Status:day 2	Cell line:H1 | Status:day 3	Cell line:H1 | Status:day 4	Cell line:H1 | Status:day 3	Cell line:H1 | Status:day 3	Cell line:H1 | Status:day 4	Cell line:H1 | Status:day 4	Cell line:H1 | Status:day 5	Cell line:H1 | Status:day 5	Cell line:H9 | Status:naive	Cell line:H9 | Status:naive	Cell line:H9 | Status:naive	Cell line:H9 | Status:primed	Cell line:H9 | Status:primed	Cell line:H9 | Status:primed	Cell line:H1 | Status:day 1	Cell line:H1 | Status:day 2	Cell line:H1 | Status:day 3	Cell line:H1 | Status:day 4	Cell line:H1 | Status:day 3	Cell line:H1 | Status:day 3	Cell line:H1 | Status:day 4	Cell line:H1 | Status:day 4	Cell line:H1 | Status:day 5	Cell line:H1 | Status:day 5	Cell line:H9 | Status:naive	Cell line:H9 | Status:naive	Cell line:H9 | Status:naive	Cell line:H9 | Status:primed	Cell line:H9 | Status:primed	Cell line:H9 | Status:primed
Pyruvate	19130.54	13694.65	15617.58	9440.81	14266.6	182114.83	153551.5	175914.45	142518.55	183612.69	163362.08	204710.05	207015.23	270089.25	204316.86	103718.54	189152.64	138419.66	99691.62	139186.45	115717.85	145343.23	127989.41	181817.05	180480.73	173681.59	168964.22	218358.3	203979.38	274755.53	237808.42	260657.75	290514.28	228701.52	225358.33	190077.12	195382.58
L-Alanine	2450.7	3066.22	3293.82	4731.79	2575.05	339421.34	236446.05	519150.84	473889.38	780970	903638.5	1470821.12	1370521.62	1947808.5	1957785.38	2113074.75	1946130.12	2243909.25	374749.91	357588.28	472814.38	47140.23	73707.6	106757.62	73674.2	76306.73	82949.48	91190.56	101064.65	92815.15	79686.85	67422.3	79448.29	56659.71	86660.25	84800.91	75427.91
lactate	35038.45	34709.1	54222.32	38128.69	59305.89	39271008	35160632	16831590	18496208	13251512	13463163	12882235	13242920	9562433	9238140	80045408	75418624	70192648	63984236	73691592	74447072	48745.69	45999.29	61669.03	42306.02	29415.42	34979.46	31395.12	31369.03	27680.12	25902	29108.72	32085.23	21361.55	60644.89	42148.88	70452.95
Acetoacetate	3908.11	7455.8	9128.84	3862.59	5712.08	1615662	1348549.5	1963866.38	1571996	1959971.12	1897742.88	2589767.25	1930904.62	3137717.75	3211359.25	1639112.5	1783902.38	1759722.12	764795.56	1007625.69	1149230	1660988	1512716	2075122.88	1671743.88	2218103.25	1839119.38	2642084	2324332.25	2498177.25	2327350.25	2935162.75	3037026	3014187	2088448	1949001	2153431.75
L-Serine	-	-	-	-	-	289708.41	275413.69	329299.5	290979.53	360013.34	369005.59	500931.41	450067.09	628127.81	545649.75	389498.38	377351.25	360384.5	256388.12	221676.25	298540.16	281464.78	295415.91	380613.75	336548.16	366503.5	342688.88	476457.03	544730.38	585021.19	492475.38	488789.16	546507.19	434227.38	329315	331335.31	380855.91
MALEAMATE	-	-	-	1200.14	-	-	-	-	-	-	-	-	3900.12	6842.18	-	8989.46	-	5165.68	21590.35	39417.12	25662.87	3057.55	4617.41	-	1850.17	-	-	-	-	4687.91	-	46883.76	65169.69	63863.64	52914.14	57036.11	56540.7
L-PROLINE	1760.52	2414.14	2111.47	1633.67	1957.94	444008.62	373576.88	363436.16	250778.61	296779.38	348126.12	279710.59	396049.97	373744.78	322141.34	413968.53	443536.16	463444.84	479165.66	524793.94	534569.38	287798.62	239950.81	338303.09	301440.84	295025	253013.38	337405.47	360030.91	304356.84	260651.23	275052.53	318393.16	263837.53	323910.28	314441.12	406261.34
3-Methyl-2-oxobutanoic acid	130635.37	110114.27	121636.31	166067.42	129996.62	578685.12	515588.03	333186.72	285483.16	262138.62	234617.19	188793.19	165824.58	153209.52	160398.66	539743.88	516072.03	484413.16	665540.69	602439.5	627783.06	131475.08	120442.51	152793.52	141153.3	102738.41	125126.25	146808.92	116460.38	145305.27	176199.02	125508.5	117481.52	134379.48	133460.73	144632.08	134640.52
Fumarate	155231.58	178497.58	143351.28	148157.8	166806.48	45667.03	57368.36	41439.5	42905.88	47972.18	45563.57	43410.61	40748.93	38510.64	58504.75	48289.69	58613.14	60212.42	54436.55	73457.59	58768.08	52754.71	54373.72	46544.36	49256.23	59138.2	77863.44	30515.99	53788.67	63658.14	63359.45	48943.82	26202.96	51308.54	47956.86	83758.02	39114.39
L-Valine	4170.12	6956.04	6295.66	4456.77	7272.61	859716	1054967.88	952495.19	1060229.5	1310949	1627942.38	1742398	1678923.5	2317524.25	2192872	2033723.62	1811228.38	2086597.12	1010601.31	873803.81	1040039.19	879564.06	911877.81	1297077	1190414	1371562.12	1315609.12	1784044.38	1842970.5	1858422	1723391.62	1769700	1794101.62	1979473	1270628	1159016	1130607.62
Succinate	588461.25	554479.25	591843.81	634912.69	683580.31	228274.3	198275.83	152032.64	148488.92	157764.3	183955.8	229268.64	216194.62	345259.22	282181.16	741049.69	681969.06	659760.69	271586.94	223916.27	316427.5	136527.02	135451.33	153265.72	158450.75	157331.22	147371.31	117852.52	136256.09	128514.4	169901.27	146866.94	151103.58	118367.71	137249.03	149942.98	110164.21
L-Threonine	-	-	-	-	-	642443.44	647389.31	794553.75	635429.25	957953.31	991489.94	1043902.44	1252785.38	1681741	1410001.38	1219223.62	1339526.75	1230239	585445.94	541132.56	718865.44	575528.19	525166.62	875582.69	720344.94	926060.81	720643.94	1255015.38	1328702.75	1443650.12	1138064.12	1178013.5	1354626.12	1280372	770240.31	713497.69	703267.81
4-HYDROXYBENZALDEHYDE	12120341	12338707	12830395	13172663	14162775	13499464	13108793	14241696	14286904	13914341	13257128	13689539	13524573	14883825	13390832	12405696	12139328	11430955	11671561	11922563	13658455	12699344	12555863	14043607	13622575	13989115	13894960	13784745	14522739	13845429	13534291	11942657	11649272	12488387	12788421	13641755	14212283
Thymine	3369	-	-	1791.03	1788.13	583644.75	607260.38	69761.93	429186.38	289642.84	283436.88	119304.37	133684.22	11577.96	6122.12	4219.23	11043.99	12551.91	530139	508099.25	724422.06	-	506250.25	131209.7	509322.09	297493.34	292462.88	173935.69	138715.23	2302.04	4948.16	-	-	1105.43	590889.19	589425.69	611071.81
Pyroglutamic acid	2721.91	1600.42	4558.16	2062.91	4857.51	8991.94	16827.89	11279.57	14374.27	9955.89	11476.03	12263.85	7064.14	11516.16	12355.67	21387.01	25073.22	17206.56	12488.62	23841.8	20555.47	15572.47	16403.72	10329.21	10238.67	8005.33	10988.57	12539.84	4665.63	8170.96	8946.9	2342.03	5324.6	2555.92	16662.88	18981.89	14629.24
5-Oxoproline	2279.23	2697.55	4935.87	2300.71	5141.81	165360.61	164433.08	95637.29	97324.23	82619.04	90614.6	85132.52	81205.66	90781.37	84082.52	361736.25	330745.47	302744.59	215545.44	243734.56	238194.92	84133.88	77977.8	65388.78	64031.54	61305.64	56948.7	75506.84	50612.06	60988.22	45390.56	61950.74	74593.43	74404.28	95878.31	82479.35	92377.37
Citraconic acid	126157.5	125327.9	143049.16	135090.06	135094.45	109904.97	160726.75	153945.23	137582.16	163494.67	176058.62	216717.67	194295.39	315624.06	316115.12	263399.66	281743.59	284503.41	128148.22	194243.27	137253.55	189206.89	137432.58	134819.86	166869.12	206342.06	196688.56	226752.73	228735.14	301750.22	315966.72	333727.38	307520.75	300410.53	120409.9	237570.33	137085.59
L-Leucine	22788.87	27410.22	28110.35	22137.62	20730	3907600.25	3292977.25	4464197.5	4419452	5690724.5	5933530.5	7136097.5	7047806.5	9739145	9656058	8958039	8771198	7914408.5	3648967.75	4182994.75	3863053.25	4464524.5	4364420.5	5238214.5	5566841.5	6226406.5	6175772	8330020	7125594.5	8103873.5	7754442.5	8154158.5	8137057.5	8086156.5	5236612.5	4714482	4741368.5
L-Asparagine	2328.29	2012.68	-	2294.61	2002.48	284950.72	251937.2	270066.66	210042.39	228484.42	247931.05	217860.02	253162.33	293034.19	282689.5	338935.75	371240.22	304394.97	286679.34	249174.23	332112.59	203890.27	202035.31	239558.88	192488.3	241143.52	173546.06	262811.28	257716.81	240858.31	214019.62	213889.5	218511.8	216647.98	274230.22	221479.2	230654.88
L-ORNITHINE	-	1300.18	2135.8	-	-	81089.15	86798.3	54944.57	72116.3	73741.69	55272.69	70292.91	70789.48	84420.59	68059.34	68226.81	-	52379.98	88106.09	84652.22	95739.75	54419.34	55020.22	54562.27	60818.09	62439.77	57651.97	73298.24	57561.68	57592.88	62243.34	53204.34	55672.58	49972.29	52644.57	49100.59	59674.06
Aspartate	-	2016.62	2025.64	-	-	350842.16	348376.66	267880.09	237080.38	268423.09	254729.48	242683	212994.77	280348.47	225220.08	6352.02	5248.07	5692.02	400749.78	424217.25	443383.09	343320.38	329101.09	268446.16	288847.75	295190.59	297889.5	349199.78	227065.27	276014.25	272578.56	282622.5	292497.28	335735.31	466655.38	354328.41	412604.78
malate	73831.29	54473.78	75204.91	76853.24	60526.93	279256.94	269933.5	159854.88	150569.17	139375.92	99772.63	115792.05	111099.78	73135.44	104770.29	463906.78	408498.75	351272.84	311322.84	305654.09	329612.16	82779.98	63225.16	112557.54	74500.06	95258.72	85151.05	70157.07	72001.55	86618.6	78552.92	85147.57	71979.91	83586.94	86045.06	73687.32	89975.37
Hypoxanthine	-	-	-	-	-	49933.64	66239.38	55692.74	56529.19	50051.97	49480.44	52261.52	50534.45	69465.09	67296.71	2578.69	2989.32	-	13659.71	16747.14	19357.93	79111.69	95946.6	63244.7	80820.12	75321.59	82894.98	90466.6	69434.56	62624.11	59856.84	54618.19	54776.25	49279.8	121752.84	110946.5	112827.06
a-ketoglutarate	66472.27	83542.57	70899.87	76586.8	77362.57	304476.31	95719.4	419065.12	385510.41	744191	626414.31	1283960	1325876.38	2108537.25	1902268.88	4370222	4358241.5	4169670.75	92627.69	110402.71	118901.12	75435.04	70442.02	148723.94	148668.34	219014.06	228684.88	416974.84	380866.84	417328.09	441776.16	602261.19	601808.94	558794.69	85523.28	66322.54	71416.73
L-Glutamine	-	-	-	-	-	4323567.5	4469475.5	5799356	5658594	6525652	6094453.5	7753528	7543737.5	8160076.5	7303309.5	5385495.5	5082175.5	5155346.5	3564248.25	3643408.75	4003424	5981808.5	5963541.5	4970694	4557390.5	4284458.5	3693598.75	3762125.75	3693262.75	2618275.25	2132839.75	1948623.38	2118025	1973651.12	6150644	5996224	5968493.5
L-LYSINE	-	-	-	-	-	158099.89	138373.73	175342.08	185701.61	200981.45	237968.25	271787.53	289251.12	408667	393733.38	287043.16	261046.8	280536.59	98017.68	126136.41	135033.55	140740.17	115632.85	197548.2	220369.25	228851.08	228215.42	314148.78	285481.03	427815.5	300079.41	311433.19	241840.92	230745.83	135845.3	153579.16	145787.05
L-Glutamate	-	-	-	-	-	1281139.88	1296751.62	738412.5	772735.94	649432.38	633733.75	666581.12	661634.19	840398.31	705659.5	2324368	2211544.75	1957706.62	1365826.25	1443171.5	1413685.62	507063.25	512407.62	499872.25	471684.16	561597	417668.25	565885.69	423865.84	425985.5	421246.12	356678.09	429489.75	440719.03	638680.88	553971.06	625487.44
CITRAMALATE	45603.74	47739.92	55443.51	43812.34	40555.04	71316.07	54732.2	173086.38	155317.27	418849.75	353425.66	611960.25	502383.16	897243.81	914823.81	752391.75	868724.19	693905.88	67507.51	58699.31	84642.1	57547.12	41442.65	213646.61	193688.27	417729.34	458092.97	795963	603164.31	837821.19	780904.56	851473.94	917998.19	1007777.81	69444.34	55919.21	57560.39
L-METHIONINE	-	-	-	-	-	286431.78	275194.97	335814.28	292412.72	384550.38	348618.91	476080.5	446950.12	689392.81	515463.91	730902.31	685764.69	602135.5	250155.23	263911.34	264803.75	263541.53	257566.2	389739.97	297395.19	420764.66	375177.84	524215.03	482499.66	487789.91	441729.09	569597.06	582131.81	599371.81	421980.53	359714.59	329722.78
D-Ribose	-	-	2053.01	1993.34	-	315918.22	257606	295006.59	346761.19	301888.91	390638.09	361411.66	473296.59	496896.16	537915.5	367687.5	326818.16	348849.09	193645.67	213549.95	211118.06	269555.5	300980.84	349158.28	292936.97	333257.28	358308.03	516141.72	460834.84	537534.75	409659.25	508513.59	601256	481785.75	342813.41	347406.16	391417.91
L-HISTIDINE	-	-	1534.01	-	-	149887.89	134753.44	166616.41	122165.65	178976.62	166934.58	187545.89	181554.98	248153.3	181314.45	295310.12	265208.03	236951.95	143156.45	194678	150681.39	153243.95	124527.94	157272.56	142177.22	174409.58	135323.7	196417.23	137475.81	197155.14	156308.81	231407.39	253607.73	235015.75	188961.8	171120.7	173221.95
3-HYDROXY-3-METHYLGLUTARATE	4078.8	6469.26	3983.17	7980.8	6746.25	2230748.75	1927511.62	2690037.75	2333885.25	2896827.25	2723695	3466655.25	3154147.25	4732692	4305195	2600344.25	2543730.25	2205262.75	1214470.5	1535843	1478572.5	2540090.25	2153908.25	2833253.75	2006453.38	2875444	2635042.5	3801537.25	3686770	3970222.75	3670132.75	4489255.5	4617478.5	5054112.5	2874982.25	2360998.75	3103778
L-Phenylalanine	1639.75	1883.46	1814.51	1814.96	-	396787.75	314586.5	385426.91	362564.34	501689.03	477918.34	577197.19	559066.44	867011.94	739810.19	842985.19	690500.75	739564.25	347526.72	429783	405396.5	320121.5	329886.66	419875.5	385798.88	427455	496825.47	632318.06	542157.31	611350.25	588478.25	612752.31	675043.31	740822.25	465877.75	435635.59	403625.16
arginine	-	1668.73	-	-	-	253633	235707.92	316862.97	333160.56	326989.34	347919.5	339956.47	325098.16	469464.12	447457.75	351328.03	365049.91	301246.84	190545.11	208534.3	206611.95	325605.44	323139.28	359118.97	374372.66	396915.34	267576.84	384776.12	359057.38	432589.53	342248.91	419019.47	410833.47	367840.22	402630.09	367351.16	319790.97
N-ACETYL-L-ASPARTIC ACID	5110.1	4589.26	3517.8	-	3764.54	46495.58	16362.37	15478.82	24259.91	-	6558.8	10655.89	7526.47	5605.69	-	85398.35	97948.73	98853.51	174206.17	159901.56	197749.7	-	-	-	-	-	10819.21	-	-	-	3469.08	-	-	-	5273.05	-	5749.66
D-Glucono-1-5-lactone	-	-	-	-	-	20648.9	22559.76	18096.56	23677.26	22488.88	25364.82	31531.74	21414.21	30428.28	29071.79	17490.29	15615.14	19837.64	15548.11	20724.13	11070.87	17163.64	19440.32	21695.1	17551.29	8451.03	24927.88	28912.7	31313.24	24769.1	34573.23	19130.63	36592.55	39743.73	25934.31	35077.39	22569.24
ALPHA-D-GLUCOSE	-	1933.92	-	-	1735.56	10349601	13612131	12783976	15188419	13236288	16036875	18604634	19028898	21679874	20869592	16209597	15258132	15159179	9211338	10893488	10239907	12456599	11192485	15794752	13965488	15497208	16218352	17058792	19182670	20568738	17738600	20077534	22175502	19670502	15280016	13944167	14190363
L-Tyrosine	-	-	3665.55	-	-	695551.25	595423.44	694248.5	714122.81	845073.19	825113.75	975179.06	1029464	1390664	1368573.38	1303688.25	1201992.12	1030590.69	637179.06	706227.19	682411.69	611622.44	568244.88	633757.19	732353.5	855718.44	869864.25	1231342.88	1024707.19	1003987.69	1066765.88	1044744.31	974649.19	1013821.69	773290.31	703764.31	726129.75
O-Phospho-L-serine	-	-	-	-	-	27821.19	35889.78	20870.69	15698.53	12404.66	17958.59	19522.15	18376.12	27605.63	19563.23	50901.29	48735.73	45859.63	33260.41	40221.35	40355.99	16871.01	13394.25	9838.24	16616.81	20643.8	11625.25	17630.64	11612.83	13095.15	11974.62	6347.02	5463.72	14578.23	17712.24	23395.95	15202.89
citrate	49358.65	45673.14	80350.16	55952.56	47513.83	56145.2	45575.93	67880.01	39623.57	70259.88	53802.95	75142.93	54032.04	48181.32	54844.17	61397.78	69019.29	66703.8	59134.64	79445	92671.48	70997.75	44674.29	72992.62	50037.54	72461.71	57079.08	63652.67	66846.04	64910.65	51807.25	63864.44	66192.84	77720.04	55658.97	90843.2	78696.92
DEOXYCYTIDINE	-	-	-	-	3257.36	11410.38	10074.96	15884.27	11169.97	25564.61	16558.79	19719.1	27202.61	27278.06	26374.87	41340.43	21263.19	-	-	-	-	12966.23	13875.99	12214.86	16783.04	15434.94	16871.85	30493.58	17204.36	22202.93	23210.86	-	-	3788.9	1787.77	-	-
deoxyuridine	1480.29	-	-	-	-	1884190.62	1810675.88	2141823.75	2040774.38	2286515.75	2160404.25	2272751.75	2193513	2818057	2631911	1668453	1420365	1594501.88	1673291.88	1533058.5	1926762.62	2301301.75	2270936.75	2061378	1726835.62	1526904.88	1416457.88	1292642.25	1125106.38	808345.69	736126.69	648429.94	753916.31	596630.12	2792568	2668845.75	2781867
Cystine	-	-	-	-	-	87606.66	87359.57	96976.23	96630.65	113112.72	124580.32	162435.11	116875.21	140273.98	191362.83	52296.25	34932.07	47853.09	98041.8	86392.07	109891.41	134472.48	103330.98	144953.78	115764.41	112191.56	144840.77	142842.77	163883.94	150971.19	134175.2	155008.77	136900.28	164882.42	150323.02	144961.61	120719.3
thymidine	2099.23	1402.95	-	-	-	17692.1	31555.21	27591.18	25143.54	35464.19	34737.07	49843.46	42311.27	36777.55	42841.08	35096.76	35842.25	28790.62	27798.8	39519.33	31028.43	22799.96	27294.02	27119.82	14644.48	33440.7	41491.48	41356.04	47035.8	39929.65	53738.59	45567.09	69687.25	52523.11	20620.73	39523.79	42954.32
aminoimidazole carboxamide ribonucleotide	-	1281.62	-	-	-	150325.12	175442.17	197047.81	202017.17	209112.25	206660.45	213083.52	267491.09	299594.88	314756.5	192979.69	186068.31	214110.05	212983.67	177280.17	185291.69	172661.3	172911.83	182497.86	206883.45	196894.2	226672	289777.84	280078.62	323609.22	243526.12	260285	356325.75	306157.34	218484	194619.44	180596.19
Sedoheptulose bisphosphate	-	1618.87	-	-	1628.13	204503.38	181436.52	179948.23	166163.23	149707.27	191334.05	160387.28	172255.55	141239.53	174896.14	142470.7	99946.9	133334.92	-	-	-	166259.33	150327.38	168523.61	177953.14	138601	155166.33	139415.02	135615.92	152619.38	147690.27	-	-	-	-	-	-
folate	-	-	-	-	-	49305.08	22741.35	20851.16	35534.73	56980.86	36711.23	54818.01	40864.21	58472.7	69159.59	47748.48	50329.42	45928.58	37361.75	34370.07	44450.89	40099.86	36853.76	51748	46575.79	37480.98	51061.41	46740.22	58256.88	57959.86	43316.42	42572.25	27801.05	47728.3	33395.26	35552.9	24466.04
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
Pyruvate
L-Alanine
lactate
Acetoacetate
L-Serine
MALEAMATE
L-PROLINE
3-Methyl-2-oxobutanoic acid
Fumarate
L-Valine
Succinate
L-Threonine
4-HYDROXYBENZALDEHYDE
Thymine
Pyroglutamic acid
5-Oxoproline
Citraconic acid
L-Leucine
L-Asparagine
L-ORNITHINE
Aspartate
malate
Hypoxanthine
a-ketoglutarate
L-Glutamine
L-LYSINE
L-Glutamate
CITRAMALATE
L-METHIONINE
D-Ribose
L-HISTIDINE
3-HYDROXY-3-METHYLGLUTARATE
L-Phenylalanine
arginine
N-ACETYL-L-ASPARTIC ACID
D-Glucono-1-5-lactone
ALPHA-D-GLUCOSE
L-Tyrosine
O-Phospho-L-serine
citrate
DEOXYCYTIDINE
deoxyuridine
Cystine
thymidine
aminoimidazole carboxamide ribonucleotide
Sedoheptulose bisphosphate
folate
METABOLITES_END
#END