{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002373","ANALYSIS_ID":"AN003868","VERSION":"1","CREATED_ON":"November 29, 2022, 8:18 pm"},

"PROJECT":{"PROJECT_TITLE":"Extracellular metabolome of activated CD8+ T cells","PROJECT_SUMMARY":"WT OT-I CD8+ T cells were activated with plate-bound anti-CD3 and soluble anti-CD28. Blank media or media post 10hr, 30hr or 48hr cell culture were collected for mass spectrometry analysis.","INSTITUTE":"Johns Hopkins University","LAST_NAME":"Xu","FIRST_NAME":"Wei","ADDRESS":"1650 Orleans Street, Baltimore, MD 21287, USA.","EMAIL":"wxu29@jhmi.edu","PHONE":"443-220-9936"},

"STUDY":{"STUDY_TITLE":"Extracellular metabolome of activated CD8+ T cells","STUDY_SUMMARY":"WT OT-I CD8+ T cells were activated with plate-bound anti-CD3 and soluble anti-CD28. Blank media or media post 10hr, 30hr or 48hr cell culture were collected for mass spectrometry analysis.","INSTITUTE":"Johns Hopkins University","LAST_NAME":"Xu","FIRST_NAME":"Wei","ADDRESS":"1650 Orleans Street, Baltimore, MD 21287, USA.","EMAIL":"wxu29@jhmi.edu","PHONE":"443-220-9936"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"C57BL/6J","AGE_OR_AGE_RANGE":"6-8 weeks","GENDER":"Male and female"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"10hr_M_01",
"Factors":{"Treatment":"10hr blank media"},
"Additional sample data":{"RAW_FILE_NAME":"10hr_M_01.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"10hr_M_02",
"Factors":{"Treatment":"10hr blank media"},
"Additional sample data":{"RAW_FILE_NAME":"10hr_M_02.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"10hr_M_03",
"Factors":{"Treatment":"10hr blank media"},
"Additional sample data":{"RAW_FILE_NAME":"10hr_M_03.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"10hr_WT_01",
"Factors":{"Treatment":"media post 10hr cell activation"},
"Additional sample data":{"RAW_FILE_NAME":"10hr_WT_01.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"10hr_WT_02",
"Factors":{"Treatment":"media post 10hr cell activation"},
"Additional sample data":{"RAW_FILE_NAME":"10hr_WT_02.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"10hr_WT_03",
"Factors":{"Treatment":"media post 10hr cell activation"},
"Additional sample data":{"RAW_FILE_NAME":"10hr_WT_03.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"30hr_M_01",
"Factors":{"Treatment":"30hr blank media"},
"Additional sample data":{"RAW_FILE_NAME":"30hr_M_01.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"30hr_M_02",
"Factors":{"Treatment":"30hr blank media"},
"Additional sample data":{"RAW_FILE_NAME":"30hr_M_02.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"30hr_M_03",
"Factors":{"Treatment":"30hr blank media"},
"Additional sample data":{"RAW_FILE_NAME":"30hr_M_03.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"30hr_WT_01",
"Factors":{"Treatment":"media post 30hr cell activation"},
"Additional sample data":{"RAW_FILE_NAME":"30hr_WT_01.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"30hr_WT_02",
"Factors":{"Treatment":"media post 30hr cell activation"},
"Additional sample data":{"RAW_FILE_NAME":"30hr_WT_02.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"30hr_WT_03",
"Factors":{"Treatment":"media post 30hr cell activation"},
"Additional sample data":{"RAW_FILE_NAME":"30hr_WT_03.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"48hr_M_01",
"Factors":{"Treatment":"48hr blank media"},
"Additional sample data":{"RAW_FILE_NAME":"48hr_M_01.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"48hr_M_02",
"Factors":{"Treatment":"48hr blank media"},
"Additional sample data":{"RAW_FILE_NAME":"48hr_M_02.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"48hr_M_03",
"Factors":{"Treatment":"48hr blank media"},
"Additional sample data":{"RAW_FILE_NAME":"48hr_M_03.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"48hr_WT_01",
"Factors":{"Treatment":"media post 48hr cell activation"},
"Additional sample data":{"RAW_FILE_NAME":"48hr_WT_01.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"48hr_WT_02",
"Factors":{"Treatment":"media post 48hr cell activation"},
"Additional sample data":{"RAW_FILE_NAME":"48hr_WT_02.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"48hr_WT_03",
"Factors":{"Treatment":"media post 48hr cell activation"},
"Additional sample data":{"RAW_FILE_NAME":"48hr_WT_03.wiff"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Blank media or media post cell culture were collected and centrifuged at 1200rpm for 5min. Supernatant were flash frozen in liquid nitrogen until further analysis.","SAMPLE_TYPE":"Cultured cells"},

"TREATMENT":{"TREATMENT_SUMMARY":"CD8+ T cells were isolated from spleens and lymph nodes from WT mice. Cells were activated with plate-bound anti-CD3 and soluble anti-CD28. Blank media post 10, 30, 48 hours in the incubator or media post 10, 30, 48 hr activated CD8+ T cell culture were collected for MS analysis."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"For measurement of extracellular metabolome, 625 µL of media was added to 375 µL acetonitrile ACN. Samples were stored at -20 °C for at least two hours followed by centrifugation at 14,000xg to precipitate any proteins. 250 µL of the supernatant was mixed with 250 µL of water and added to a 3 kDa molecular weight cut-off filter spin column (Microcon YM-3 Centrifugal Filter, Millipore). Samples were centrifuged at 14,000xg at 4 °C for 30 min. The flow-through was saved for LC-MS/MS analysis."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Shimadzu Prominence UFLC","COLUMN_NAME":"Waters XBridge BEH Amide, 2.1 x 150 mm, 2.5 μm"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"ABI Sciex 5500 QTrap","INSTRUMENT_TYPE":"QTRAP","MS_TYPE":"ESI","ION_MODE":"UNSPECIFIED","MS_COMMENTS":"The optimized MS parameters were: ESI voltage was +5,000 V in positive ion mode and -4,500 V in negative ion mode; dwell time was 3 ms per SRM transition and the total cycle time was 1.57 seconds. Peak integration for each targeted metabolite in SRM transition was processed with MultiQuant software (v2.1, AB Sciex). The preprocessed data with integrated peak areas were exported from MultiQuant and re-imported into Metaboanalyst software for further data analysis (statistical analysis, principal component analysis, generating heatmap, enrichment analysis, etc.)."},

"MS_METABOLITE_DATA":{
"Units":"AUC",

"Data":[{"Metabolite":"Urea","10hr_M_01":"285141357.4","10hr_M_02":"271538886.7","10hr_M_03":"289128554.3","10hr_WT_01":"272680127.8","10hr_WT_02":"278166339.8","10hr_WT_03":"289444513.9","30hr_M_01":"275536866.1","30hr_M_02":"300726825.1","30hr_M_03":"295045640.1","30hr_WT_01":"277773688.9","30hr_WT_02":"286569670.2","30hr_WT_03":"275182193.5","48hr_M_01":"297534553.1","48hr_M_02":"297780738.5","48hr_M_03":"300201644.7","48hr_WT_01":"315867195.7","48hr_WT_02":"299863924.4","48hr_WT_03":"307981930.4"},{"Metabolite":"ethanolamine","10hr_M_01":"1605893.824","10hr_M_02":"1653754.591","10hr_M_03":"1665939.495","10hr_WT_01":"1680882.794","10hr_WT_02":"1741083.853","10hr_WT_03":"1889932.917","30hr_M_01":"1772775.54","30hr_M_02":"1632436.167","30hr_M_03":"1870343.006","30hr_WT_01":"1835777.688","30hr_WT_02":"1907029.433","30hr_WT_03":"2018409.766","48hr_M_01":"1982764.57","48hr_M_02":"1872757.279","48hr_M_03":"2391871.804","48hr_WT_01":"1700321.077","48hr_WT_02":"1977921.733","48hr_WT_03":"2086763.363"},{"Metabolite":"Imidazole","10hr_M_01":"2316502.919","10hr_M_02":"2572153.409","10hr_M_03":"2734464.471","10hr_WT_01":"2957634.212","10hr_WT_02":"2763957.107","10hr_WT_03":"2673012.452","30hr_M_01":"2318772.346","30hr_M_02":"2471828.053","30hr_M_03":"2483431.333","30hr_WT_01":"2682443.599","30hr_WT_02":"2529514.476","30hr_WT_03":"1958157.83","48hr_M_01":"3078052.104","48hr_M_02":"2052492.876","48hr_M_03":"1649737.085","48hr_WT_01":"2841008.604","48hr_WT_02":"2185000.967","48hr_WT_03":"2212902.447"},{"Metabolite":"glycine","10hr_M_01":"558629.8756","10hr_M_02":"476370.3366","10hr_M_03":"454508.9693","10hr_WT_01":"598002.2086","10hr_WT_02":"443518.5256","10hr_WT_03":"478845.573","30hr_M_01":"379587.3657","30hr_M_02":"559177.4045","30hr_M_03":"466157.562","30hr_WT_01":"466327.352","30hr_WT_02":"430197.5347","30hr_WT_03":"393044.3321","48hr_M_01":"581550.3113","48hr_M_02":"490167.8247","48hr_M_03":"512803.6529","48hr_WT_01":"464070.6405","48hr_WT_02":"415660.0161","48hr_WT_03":"582477.9085"},{"Metabolite":"alanine","10hr_M_01":"26529350.76","10hr_M_02":"25218143.75","10hr_M_03":"25741884.18","10hr_WT_01":"28912024.73","10hr_WT_02":"29522626.23","10hr_WT_03":"28442508.54","30hr_M_01":"26981059.13","30hr_M_02":"25332390.24","30hr_M_03":"26457868.3","30hr_WT_01":"24168319.39","30hr_WT_02":"23695046.68","30hr_WT_03":"25324781.41","48hr_M_01":"27273021.06","48hr_M_02":"27980593.36","48hr_M_03":"29132843.84","48hr_WT_01":"31641191.51","48hr_WT_02":"31332733.12","48hr_WT_03":"31781550.4"},{"Metabolite":"4-aminobutyrate","10hr_M_01":"152901.8896","10hr_M_02":"183871.8178","10hr_M_03":"151751.9367","10hr_WT_01":"75682.94474","10hr_WT_02":"158378.1872","10hr_WT_03":"143888.1786","30hr_M_01":"234500.7649","30hr_M_02":"180570.5453","30hr_M_03":"194827.7811","30hr_WT_01":"193452.282","30hr_WT_02":"198783.3596","30hr_WT_03":"145700.2365","48hr_M_01":"290627.8208","48hr_M_02":"102821.6143","48hr_M_03":"204214.7544","48hr_WT_01":"251817.3381","48hr_WT_02":"284150.984","48hr_WT_03":"244710.7599"},{"Metabolite":"serine","10hr_M_01":"150373322.7","10hr_M_02":"141280626.5","10hr_M_03":"138266533.9","10hr_WT_01":"145307782.2","10hr_WT_02":"144947162.1","10hr_WT_03":"151413724.7","30hr_M_01":"145364279","30hr_M_02":"145459110.1","30hr_M_03":"144406172.2","30hr_WT_01":"137331270.2","30hr_WT_02":"126843664.9","30hr_WT_03":"129888582.8","48hr_M_01":"153498508.4","48hr_M_02":"148915049","48hr_M_03":"147104554","48hr_WT_01":"100297866.9","48hr_WT_02":"95719843.68","48hr_WT_03":"97864842.15"},{"Metabolite":"cytosine","10hr_M_01":"170342.6122","10hr_M_02":"217091.2329","10hr_M_03":"182418.4958","10hr_WT_01":"219492.1521","10hr_WT_02":"233209.7624","10hr_WT_03":"208084.7736","30hr_M_01":"136187.3849","30hr_M_02":"141844.5869","30hr_M_03":"149449.0551","30hr_WT_01":"466367.9588","30hr_WT_02":"566863.7163","30hr_WT_03":"436279.344","48hr_M_01":"180659.7572","48hr_M_02":"184481.1565","48hr_M_03":"171335.1892","48hr_WT_01":"1339881.503","48hr_WT_02":"1265514.095","48hr_WT_03":"1504647.706"},{"Metabolite":"creatinine","10hr_M_01":"105803687.3","10hr_M_02":"100801059.5","10hr_M_03":"104148866.3","10hr_WT_01":"126419862.7","10hr_WT_02":"130183454.5","10hr_WT_03":"126881550.5","30hr_M_01":"99548623.5","30hr_M_02":"106872310","30hr_M_03":"109080541","30hr_WT_01":"108209219.9","30hr_WT_02":"108430529","30hr_WT_03":"108041543.1","48hr_M_01":"109860738.2","48hr_M_02":"115706952.1","48hr_M_03":"114943853.4","48hr_WT_01":"108961669.5","48hr_WT_02":"112102992.5","48hr_WT_03":"120919691"},{"Metabolite":"proline","10hr_M_01":"52307702.64","10hr_M_02":"51115220.64","10hr_M_03":"49224314.9","10hr_WT_01":"55236982.43","10hr_WT_02":"52513589.3","10hr_WT_03":"50090415.42","30hr_M_01":"53986127.73","30hr_M_02":"50428776.44","30hr_M_03":"51744454.71","30hr_WT_01":"44980164.32","30hr_WT_02":"45997684.43","30hr_WT_03":"50926154.9","48hr_M_01":"51339204.94","48hr_M_02":"50883426","48hr_M_03":"54871703.27","48hr_WT_01":"46299502.45","48hr_WT_02":"48784632.26","48hr_WT_03":"436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}

}