Blood glucose was determined using an Accu-Chek Aviva system (Roche Diagnostics, Risch, Switzerland). Commercial kits for measurement of serum insulin (Mercodia, Uppsala, Sweden) and non-esterified fatty acids (NEFA; Wako Chemicals GmbH, Neuss, Germany) were applied following the manufacture’s protocols. Indexes of insulin resistance and sensitivity were derived from glucose, insulin and NEFA levels in plasma prepared from the blood of 6 h-fasted animals. The homeostatic model assessment for insulin resistance (HOMA-IR) score was calculated as HOMA-IR = [insulin (μU/L) × glucose (mmol/L)/22.5] [29 (link)], and the revised quantitative insulin sensitivity check index (R-QUICKI) as R-QUICKI = 1/[log glucose (mg/dL) + log insulin (μU/mL) + log NEFA (mM/L)] [30 (link)].
Accu chek aviva system
Manufactured by Roche
Sourced in Switzerland
The Accu-Chek Aviva system is a blood glucose monitoring device designed to help individuals manage their diabetes. It provides blood glucose readings to assist in monitoring and managing blood sugar levels.
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Lab products found in correlation
Mouse insulin elisa kit, by Merck Group (2 mentions)
Sterile saline, by Merck Group (2 mentions)
Thrombin, by Merck Group (1 mentions)
Fibrinogen from human plasma, by Merck Group (1 mentions)
Streptozotocin (stz), by Merck Group (1 mentions)
Streptozotocin (stz), by Thermo Fisher Scientific (1 mentions)
Edta treated tubes, by Sarstedt (1 mentions)
Reflotron plus system, by Roche (1 mentions)
10 protocols using accu chek aviva system
1
Insulin Resistance Biomarkers in Fasted Animals
2
Plasma Insulin and Glucose Tolerance Test
3
Transplantation of Differentiated iPIC Aggregates
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Differentiated iPIC aggregates were mixed with 100 μL of fibrinogen/50 μL of thrombin solution, incubated at 37 °C for 5 min and then implanted in the subcutaneous space of anaesthetized STZ-NOD-scid mice (3–4 × 106 cells/mouse). Fibrinogen from human plasma (Merck Millipore) and thrombin (Sigma) were reconstituted in iMEM and in PBS to make 10 mg/mL and 50 IU/mL solutions, respectively, and stored at − 80 °C until use. For the kidney capsule implantation study (Supplementary Fig. 7 ), s6-iPIC-A (1.4 × 106 cells/mouse) or s6-iPIC-B (3.6 × 106 cells/mouse) was implanted directly into the kidney capsule without fibrin gel. We monitored the blood glucose levels of implanted animals using an Accu-Chek Aviva system (Roche DC Japan) and collected plasma samples from the tail vein on the indicated days. For the oral glucose tolerance test, the mice were fasted overnight and orally injected with a 2 g/kg glucose solution (Otsuka), and plasma samples were collected from the tail vein before and 15, 30, 60 and 120 min after injection.
4
Plasma Insulin and Glucose Tolerance Test
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Plasma insulin measurement and GTT were performed at the end of phase II. After overnight fasting, blood was collected via tail tip transection and plasma was isolated by centrifugation. Fasting plasma insulin was measured using a mouse insulin ELISA kit with rat insulin as a standard (EMD Millipore, MO). Fasting blood glucose was measured using ACCU-CHEK Aviva system (Roche Diagnostics, Switzerland). Mice were then injected intraperitoneally with 20% dextrose solution in sterile saline (Sigma-Aldrich, MO) at the dosage of 0.75 g dextrose per 1 kg of body weight. Blood glucose was measured at 15, 30, 45, 60, 90, and 120 minutes after the injection. Glucose intolerance was measured by calculating the area under the curve in blood glucose vs. time graph using trapezoidal method.
5
Oral Glucose Tolerance Test in Mice
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For oGTT, mice were fasted overnight (14–18 h) and then given a glucose load (25% stock solution in saline) of 2 g per kg of body weight by oral administration. For ITT, intraperitoneal injection of an insulin bolus of 4 IU per kg of body weight was performed. Blood samples were collected from the tail vein at 0, 15, 30, 60 and 120 min after administration of glucose or insulin. Plasma glucose concentration was measured using an Accu-Chek Aviva system (Roche Diagnostics).
6
Streptozotocin-Induced Diabetes Model in Mice
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For Streptozotocin (STZ, Sigma-Aldrich, USA) treatment, 4-month-old female C57BL/6J B6 albino mice were fasted for 6 hr and a single dose of 200 mg STZ per kg body weight in phosphate buffered saline (PBS, Thermo Fisher Scientific, USA) was injected intraperitoneally. For glucose tolerance tests, animals were fasted for 6 hr and 2 g D-glucose (20% solution in PBS+/+) per kg body weight was injected intraperitoneally. Blood glucose values were measured afterwards at specified time points using an Accu-Chek Aviva system (Roche, Switzerland).
7
Diphtheria Toxin Induced Diabetes Model
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Treatment consisted of a single i.p. injection of 500 ng of diphtheria toxin in phosphate buffered saline (PBS). Blood glucose was measured using an Accu-Chek Aviva system (Roche, Bromma, Sweden). For the assessment of insulin levels, plasma was collected in EDTA-treated tubes (Sarstedt, Nümbrecht, Germany) and measured with alphaLISA (Perkin Elmer, Upplands Väsby, Sweden). Glucose tolerance tests were performed after 12 hours overnight fasting, by giving an i.p. injection of 2 g glucose per kg bodyweight. To avoid prolonged hyperglycemia, DT-treated RIP-DTR mice used for subsequent OPT imaging were transplanted with ~100–150 RIP-DTR negative islets into the ACE, normalising their blood glucose levels as shown previously for streptozotocin-treated mice43 (link).
8
Evaluating Metabolic Parameters in Mice
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Mouse body weights (BWs) were measured, and blood samples were collected from the tail vein every 15 days during the 12 weeks of nutritional intervention. Mice were fasted for 12 h before blood collection. Blood glucose concentrations were measured using an automatic glucometer (Accu-Chek Aviva System, Roche, Indianapolis, IN, USA). Serum aspartate aminotransferase (GOT/AST) activity was analyzed with a Reflotron Plus system (Roche Diagnostics, Burgess Hill, West Sussex, UK) according to the manufacturer’s protocols.
9
Transgenic Mouse Model for Glucose Metabolism
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Both transgenic (RIP-CD154 × RAG KO (n = 6) and RAG KO (n = 3) on a C57BL/6 background) and control mice (C57BL/6) (n = 10) were bred at Taconic M&B, Ry, Denmark, and genotyped by PCR as previously described (Haase et al. 2004 (link); Haase and Markholst 2007 (link)). The animals, all female and 12 weeks old, were housed in polyethylene cages in a temperature, humidity and 12 h day–night rhythm controlled room receiving standard chow. After weighing of the animals, blood samples were collected retro-orbital, without anesthesia, into eppendorf tubes and the animals sacrificed immediately afterwards. The blood samples were left for coagulation and hereafter centrifuged (15 min at 2000×g at RT) for the collection of serum. Blood glucose was measured in serum using the Accu-Chek Aviva system by Roche. All animal experiments were conducted according to Danish legislation and approved by the Danish Animal Inspectorate.
10
Biomarkers of Insulin Sensitivity
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Blood glucose was determined using an Accu-Chek Aviva system (Roche Diagnostics, Risch, Switzerland). Commercial kits for measurement of serum insulin (Mercodia, Uppsala, Sweden), non-esterified fatty acids (NEFA; Wako Chemicals GmbH, Neuss, Germany) and triacylglycerides (TAG; Sigma-Aldrich, St. Louis, MO, USA) were applied following the manufacturer’s protocols. The homeostatic model assessment for insulin resistance (HOMA-IR) and the revised quantitative insulin sensitivity check index (R-QUICKI) were calculated as described earlier [28 (link)].
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