{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002960","ANALYSIS_ID":"AN004861","VERSION":"1","CREATED_ON":"November 3, 2023, 5:49 am"},

"PROJECT":{"PROJECT_TITLE":"Mid-Old Cells are Potential Target for Anti-aging Interventions in the Elderly","PROJECT_SUMMARY":"The biological process of aging is thought to result in part from accumulation of senescent cells in organs. However, the present study identified a subset of fibroblasts and smooth muscle cells which are the major constituents of organ stroma neither proliferative nor senescent in tissues of the elderly, which we termed “mid-old status” cells. Upregulation of pro-inflammatory genes (IL1B and SAA1) and downregulation of anti-inflammatory genes (SLIT2 and CXCL12) were detected in mid-old cells. In the stroma, SAA1 promotes development of the inflammatory microenvironment via upregulation of MMP9, which decreases the stability of epithelial cells present on the basement membrane, decreasing epithelial cell function. Remarkably, the microenvironmental change and the functional decline of mid-old cells could be reversed by a young cell-originated protein, SLIT2. Our data identify functional reversion of mid-old cells as a potential method to prevent or ameliorate aspects of aging-related tissue dysfunction.","INSTITUTE":"Ajou University Medical Center","LAST_NAME":"Kim","FIRST_NAME":"Young Hwa","ADDRESS":"206, World cup-ro, Yeongtong-gu, Suwon-si, Gyeonggi-do, Republic of Korea","EMAIL":"skyblue32@nate.com","PHONE":"+82-10-5153-3636"},

"STUDY":{"STUDY_TITLE":"Analysis of Lipids Secreted from Fibroblast Young Cells","STUDY_SUMMARY":"In this experimental study, we aimed to understand the potential factors within the secretions of young cells that could trigger the reverse aging of Mid-old cells. To investigate this phenomenon, we co-cultured young cells with Mid-old cells and observed a fascinating outcome: the Mid-old cells exhibited reverse aging and transformed into a more youthful state. To uncover the specific factors responsible for this reverse aging effect, we conducted a detailed analysis of the secreted factors from the young cells. Our analysis focused on a range of biomolecules, including lipids. However, despite our efforts, we did not identify any distinct factors that could be directly attributed to this remarkable reverse aging process.","INSTITUTE":"Ajou University Medical Center","LAST_NAME":"Kim","FIRST_NAME":"Young Hwa","ADDRESS":"206, World cup-ro, Yeongtong-gu, Suwon-si, Gyeonggi-do, Republic of Korea","EMAIL":"skyblue32@nate.com","PHONE":"+82-10-5153-3636"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"20210625_AJU_Y1_lipidomics",
"Factors":{"*Factor":"Test sample"},
"Additional sample data":{"RAW_FILE_NAME":"20210625_AJU_Y1_lipidomics.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"20210625_AJU_Y2_lipidomics",
"Factors":{"*Factor":"Test sample"},
"Additional sample data":{"RAW_FILE_NAME":"20210625_AJU_Y2_lipidomics.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"20210625_AJU_Y3_lipidomics",
"Factors":{"*Factor":"Test sample"},
"Additional sample data":{"RAW_FILE_NAME":"20210625_AJU_Y3_lipidomics.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"20210625_AJU_M1_lipidomics",
"Factors":{"*Factor":"Control sample"},
"Additional sample data":{"RAW_FILE_NAME":"20210625_AJU_M1_lipidomics.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"20210625_AJU_M2_lipidomics",
"Factors":{"*Factor":"Control sample"},
"Additional sample data":{"RAW_FILE_NAME":"20210625_AJU_M2_lipidomics.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"20210625_AJU_M3_lipidomics",
"Factors":{"*Factor":"Control sample"},
"Additional sample data":{"RAW_FILE_NAME":"20210625_AJU_M3_lipidomics.mzML"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"The experiment was initiated by seeding the cells in a 100mm cell culture dish, followed by a 2-day incubation period, during which the culture medium was carefully harvested from the cell culture dish. Use a sterile technique to minimize contamination. Transfer the harvested culture medium to a suitable container.","SAMPLE_TYPE":"Cultured fibroblasts"},

"TREATMENT":{"TREATMENT_SUMMARY":"no treatment"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"For lipid extraction from samples, a two-step method involving neutral and acidic extraction were used. At first, in neutral extraction, lipids from the samples were extracted according to the Folch method using a mixture of chloroform and methanol (2:1, v/v). The samples were vortexed and incubated on ice for 10 min. The samples were centrifuged at 13,800×g for 2 min at 4°C, supernatant was transferred to a new 1.5 mL tube. Next, in Acidic extraction, 750 μL of chloroform:methanol:HCl (1N, 37%) (40:80:1, v/v/v) was added to the remaining samples. After incubating for 15 min at room temperature, 250 μL of cold chloroform and 450 μL of cold 0.1 M HCl were added to the sample. The mixture was vortexed for 1 min and centrifuged at 6,500×g for 2 min at 4°C. The lower organic phase was collected and combined with the prior extract. The sample was then dried using HyperVAC-MAX VC2200 centrifugal vacuum concentrator (Hanil Scientific Inc., Korea). Dried metabolite contents were reconstituted in 50 µL of mobile phase solvent A:solvent B (2 :1, v/v) and then subjected to LC-MS/MS analysis."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Q Exactive™ Hybrid Quadrupole-Orbitrap MS coupled to a 1290 Infinity UHPLC","COLUMN_NAME":"e Hypersil GOLD™ C18 HPLC column(2.1 × 100 mm; 1.9 μm particle size; Thermo Fischer Scientific)","SOLVENT_A":"Acetonitrile:Methanol:Water (19:19:2, v/v/v) + 20 mmol/L ammonium formate + 0.1% formic acid (v/v)","SOLVENT_B":"2-propanol + 20 mmol/L ammonium formate + 0.1% formic acid (v/v)","FLOW_GRADIENT":"0–5 min, 5% B; 5–15 min, 5–30% B; 15–22 min, 30–90% B; 22–25 min, 90% B; 25–26 min, 90–5% B; 26–30 min, 5% B. Parameters were set as follows: positive mode, spray voltage; 3.8 kV","FLOW_RATE":"0.2 mL/min","COLUMN_TEMPERATURE":"320℃"},

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

"MS":{"INSTRUMENT_NAME":"Q Exactive™ Hybrid Quadrupole-Orbitrap MS","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Obtained UHPLC-Orbitrap-MS/MS RAW files were processed using Lipid Search 4.2TM (Thermo Fisher Scientific, Waltham, MA, USA). Lipid profiling under the following conditions: parent search; 0.2 Da, product search; 5.0 ppm, precursor ion mass tolerance; 8.0 ppm, M-score threshold; 2.0.","MS_RESULTS_FILE":"ST002960_AN004861_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes"}

}