Serum samples were obtained from peripheral blood by centrifugation and stored at –80°C until used. Using commercially available enzyme-linked immunosorbent assays (ELISA), we measured serum concentrations of vaspin (Cat. No. RD101097200R, detection limit: 0.01 ng/ml, BioVendor, Czech Republic), visfatin (Cat. No. RAG004R, detection limit: 30 pg/ml, BioVendor, Czech Republic), irisin (Cat. No. RAG018R, detection limit: 1 ng/ml, BioVendor, Czech Republic), betatrophin (Cat. No. SK00528-06, detection limit: 160 pg/ml, Aviscera Bioscience Inc., USA), and insulin (Cat. No. ME E-0900, detection limit: 1.76 µIU/ml, LDN Labor Diagnostika Nord, Germany) in all participants. Moreover, in the patients, we measured routine blood variables such as white blood cell count (WBC), platelet count (PLT), hemoglobin (HGB) concentration; activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGTP), and alkaline phosphatase (ALP); concentrations of fasting glucose, urea, creatinine, bilirubin, total cholesterol (TCh), triglycerides (TG), and high-density lipoprotein cholesterol (HDL-Ch); prothrombin activity index (PT%); albumin and total protein concentration; and cancer markers (α-fetoprotein [AFP], carcinoembryonic antigen [CEA], cancer antigen 19-9 [CA 19-9]).
Visfatin
Manufactured by BioVendor
Sourced in Czechia
Visfatin is a protein that plays a role in various biological processes. It is a multifunctional enzyme involved in the regulation of cellular metabolism and energy production. Visfatin is expressed in a variety of tissues and has been studied for its potential involvement in various physiological and pathological conditions.
Automatically generated - may contain errors
2 protocols using visfatin
1
Serum Biomarker Measurements in Patient Cohort
2
Osteogenic Differentiation of hMSCs
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Primary hMSCs (n ¼ 4) or commercially obtained donor MSCs (n ¼ 3) were cultured at 4 Â 10 4 cells per 9.6 cm 2 in alphaMEM Glutamax expansion medium supplemented with 10% FCS (Sig-maeAldrich), 100 U/ml penicillin (PAA), 10 mg/ml streptomycin (PAA) and osteogenic supplements, 0.1 mM dexamethasone (SigmaeAldrich), 10 mM b-glycerophosphate (SigmaeAldrich), and 0.06 mM ascorbic acid (SigmaeAldrich) for 21 days 23, 25 . Cells were permanently stimulated with/without visfatin (100 ng/ml, Biovendor, Brno, Czech Republic), resistin (20 ng/ml, PeproTech, Rocky Hill, USA), and leptin (10 ng/ml, Biovendor) during differentiation. Untreated cells served as control. Culture was performed at 37 C with 5% CO 2 and medium changed every 2 days. Cells were lysed (RLT lysis buffer. Qiagen, Venlo, Netherlands) for ribonucleic acid (RNA) isolation and supernatants collected.
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