Heparin

Manufactured by Nipro

Sourced in Japan

Heparin is an anticoagulant medication used to prevent and treat blood clots. It is a naturally occurring substance found in the body that helps regulate blood clotting. Heparin is commonly used in medical settings, such as hospitals and laboratories, to maintain the fluidity of blood samples during various testing and analysis procedures.

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Lab products found in correlation

Red blood cell lysis buffer, by Merck Group (1 mentions) Versalyse, by Beckman Coulter (1 mentions) Dulbecco s modified eagle, by Thermo Fisher Scientific (1 mentions) Meloxicam, by Boehringer Ingelheim (1 mentions) L 15 medium, by Thermo Fisher Scientific (1 mentions)

5 protocols using heparin

Exhaustive Exercise-Induced Physiological Changes

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Ten animals from each group were sacrificed immediately after exhaustive exercise, and 10 at 24 h after exhaustive exercise. Anesthesia was induced with 2% isoflurane inhalation at 0.8 L·min⁻¹, maintained at 1% at 0.8 L·min⁻¹. Blood samples were collected from the abdominal aorta using a 1-mL syringe mounted with a 20-gauge needle and coated with heparin (5000 UI·mL⁻¹; Nipro, Osaka, Japan). Blood samples were transferred to a tube coated with heparin and centrifuged at 2600g for 10 min, then plasma was stored at −80°C until analysis.
The liver tissues were snap-frozen by immersing the samples in liquid nitrogen and stored at −80°C until analysis.
We isolated leukocytes from the bone marrow using previously described methods with some modifications (20 (link)). Femurs and tibiae were removed, and bone marrow was harvested by flushing with Hanks’ balanced salt solution without Ca²⁺/Mg²⁺, 30 mM HEPES, and 15 mM EDTA. A single-cell suspension was created by passing the suspension through a 21-gauge needle. The cells were centrifuged at 1200 rpm for 5 min at room temperature. The resultant pellet containing leukocytes was suspended in 5 mL of red blood cell lysis buffer (Sigma-Aldrich, St. Louis, MO) and centrifuged at 1200 rpm for 5 min at room temperature. Cells were resuspended in Versa Lyse (Beckman Coulter, Miami, FL).

MIZOKAMI T, & SUZUKI K. (2022). Neutrophil Depletion Attenuates Acute Liver Stress after Exhaustive Exercise in Mice. Medicine and Science in Sports and Exercise, 55(4), 670-679.

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Exercise-Induced Organ Sampling Protocol

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Ten animals from each group were killed immediately after exhaustive exercise, and 10 at 24 h after exhaustive exercise. Anaesthesia was induced by inhalation of isoflurane as described 2% isoflurane at 0.8 L/min until exsanguination. The depth of anaesthesia in this study was determined to be adequate based on the absence of any flexion response to a noxious stimulus, such as pinching the digit for ∼2 s. Blood samples were collected from the abdominal aorta using a 1 mL syringe mounted with a 20‐gauge needle and coated with heparin (5000 IU/mL; Nipro, Osaka, Japan). Blood samples were transferred to a tube coated with heparin and centrifuged at 2600g for 10 min, and the plasma was stored at −80°C until analysis. The kidneys were removed, and the right kidney was immersed in liquid nitrogen, snap frozen, and stored at −80°C until analysis; the left kidney was frozen in Tissue‐tek Crymould (Sakura, Torrance, CA, USA) filled with OCT compound (Sakura), with samples immersed in precooled isopentane at −80°C.

Mizokami T., Shimada M, & Suzuki K. (2024). Neutrophil depletion attenuates acute renal injury after exhaustive exercise in mice. Experimental Physiology, 109(4), 588-599.

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Beagle Bone Marrow Aspiration Protocol

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Proximal femur of healthy beagle dogs (n = 3) was used to collect the 5 mL of bone marrow samples in to 10 mL syringe containing 1 mL Dulbecco’s modified eagle’s medium (DMEM, Life technologies, New York, USA) and 1000 U/mL of heparin (Nipro, Osaka, Japan). The use of all samples from healthy experimental Beagle dogs (mean age: 12.9 months; range: 12–14 months) was in accordance with Hokkaido University Institutional Animal Care and Use Committee guidelines (approval number: 21–0022). Briefly, dogs were put under general anesthesia induced with propofol (Intervet, Tokyo, Japan) at 6 mg/kg intravenously and maintained on isoflurane (Intervet) and oxygen. Meloxicam (Boehringer-Ingelheim Animal Health, Tokyo, Japan) at 0.2 mg/kg subcutaneously was administered for pain management. The bone marrow aspiration site was aseptically prepared by clipping the hair around the proposed site of collection and scrubbed with 70% ethanol and then povidone iodine.

Wijekoon S., Sunaga T., Wang Y., Mwale C., Kim S, & Okumura M. (2022). Pentosan polysulfate regulates hepcidin 1-facilitated formation and function of osteoclast derived from canine bone marrow. PLoS ONE, 17(3), e0265596.

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Biochemical Assay Protocol

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ACh and SNP were purchased from Sigma-Aldrich (St. Louis, MO, USA). Formaldehyde, lucigenin, and phenylephrine were purchased from Nacalai Tesque Inc., Ltd., (Kyoto, Japan). Heparin was purchased from the NIPRO Corporation (Osaka, Japan).

Nakagawa K., Tanaka R., Donouchi M., Kanda M., Kamada S., Kobuchi S., Tawa M., Matsumura Y, & Ohkita M. (2022). Vascular Endothelial Dysfunction in the Thoracic Aorta of Rats with Ischemic Acute Kidney Injury: Contribution of Indoxyl Sulfate. Oxidative Medicine and Cellular Longevity, 2022, 7547269.

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Porcine DCD Liver Perfusion Model

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A DCD liver model with 60 min of circulatory arrest was constructed in pigs as reported previously.^{10 (link)} The extracted liver grafts were flushed with 500 mL of heparin saline (20 000 IU/L) and replaced with 500 mL of cold histidine-tryptophan-ketoglutarate solution. Subsequently, normothermic ex vivo perfusion was performed using our custom-made system with 2 types of perfusion solutions. For the dilution group, the perfusion solution consisted of 10% diluted donor blood, Leibovitz’s Medium (L-15 medium, Thermo fisher scientific, Waltham, MA), and pig serum (26250, Thermo fisher scientific, Waltham, MA) (total volume, 2000 mL). For the whole blood group, the perfusion solution consisted of whole blood from a donor and heparin (5000 IU/L, NIPRO, Osaka, Japan) (total volume, 1300 mL). Each experiment was conducted once.
The fluids flowing in and out of the liver were sampled every hour immediately after perfusion was initiated for metabolomic analysis. As a control group, heterotopic transplantations were also performed (in vivo group, N = 1). Under general anesthesia, blood from the portal vein (PV) and infra hepatica inferior vena cava (IH-IVC) was collected for 3 h after resuming blood flow and analyzed similar to that in the other groups (Figure 1). Metabolomic analysis was performed with 1 case in each group.

Yoshimoto S., Ohara M., Torai S., Kasamatsu H., Ishikawa J., Kimura T., Nadahara S, & Kobayashi E. (2021). Rapid Metabolic Recovery of Donor Circulatory Death Liver Graft Using Whole Blood Perfusion: A Pig Study. Transplantation Direct, 7(7), e712.

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