Contour xt

One week before the mice were sacrificed, a glucose-tolerance test was performed on mice that were food deprived for 6 h. With this aim, mice received a 50% glucose solution in water at a dose of 2 g/kg of body weight by intraperitoneal (IP) injection and then the glucose tolerance test was carried out. Tail vein blood glucose was measured just before (time 0) IP injection of glucose and 15, 30, 60 and 120 min post injection. Blood glucose levels were measured using a handheld glucometer (Contour XT, Ascensia Diabetes Care, S.L., Barcelona, Spain).

Glucose and lactate concentrations were determined in media at the beginning of and accordingly during the expansion of the cells in conditioned media by a CONTOUR^®XT (Ascensia Diabetes Care Deutschland GmbH, Leverkusen, Germany) and Lactate Plus Meter (Nova Biomedical Corporation, Waltham, MA, USA), respectively. Measured values below detection thresholds were assumed to be zero. Glucose consumption and lactate generation were calculated by the difference in respective concentrations in media and conditioned media. Both parameters were normalized to one million harvested cells and a time interval of one day. The yield of lactate from glucose was obtained by division of lactate generation by glucose consumption as described by Schop et al. [53 (link)].

During 1 week before the sacrifice, mice fasted for 8 h and were given a 2 g/kg of body weight glucose solution by intraperitoneal injection. Blood was sampled from the tail vein at 0, 15, 30, 60, and 120 min after injection. A handheld glucometer (Contour XT, Ascensia Diabetes Care, S.L., Barcelona, Spain) was used to determine glucose levels.

Blood samples were centrifuged at 1400 × g for 8 min at 4 °C. Plasma was removed and stored at 20 °C. Plasma insulin concentrations were measured with the use of enzyme-linked immunosorbent assay kits in a single assay to remove inter-assay variations (Mercodia AB, Uppsala, Sweden). Glucose levels were determined by a glucometer (Contour XT, Ascensia Diabetes Care, Milan, Italy). Basal fasting values of serum glucose and insulin were used to calculate the homoeostatic model assessment (HOMA) index as glucose (mg/dL) × insulin (mU/L)/405 [42 (link)]. Plasma concentrations of triglycerides and cholesterol, and non-esterified fatty acids (NEFAs) were measured by the colorimetric enzymatic method using commercial kits (SGM Italia, Rome, Italy and Randox Laboratories ltd., Crumlin, United Kingdom). Commercially available ELISA kits were used to determine serum adiponectin and leptin (B-Bridge International, Mountain View, CA, USA), interleukin-1α (IL-1α), interleukin-10 (IL-10), and tumor necrosis factor-α (TNF-α) (Thermo Scientific, Rockford, IL, USA; Biovendor R and D, Brno, Czech Republic).

One week before the sacrifice, mice fasted for 8 h underwent a glucose tolerance test. Glucose was administered from a 50% solution in water in a dose of 2 g/kg of body weight by intraperitoneal injection. Blood was collected at 0, 15, 30, 60, and 120 min after treatment from the tail vein and used to measure glucose levels with a handheld glucometer (Contour XT, Ascensia Diabetes Care, S.L., Barcelona, Spain).