The excised kidneys were perfused ex vivo by using the normothermic machine perfusion system consisting of the following elements: perfusion pump (Ismatec Co., Ltd., Germany), temperature control unit (Tokaihit Co., Ltd., Japan), custom-made sealed chamber for the left rat kidney (Tokaihit Co., Ltd., Japan), custom-made cannulation tube for rat kidneys (Tokaihit Co., Ltd., Japan), rodent tail vein catheter (Braintree Scientific, Inc., US), flow path tube [fluoro-rubber tube (KING WORKS Co., Ltd., Japan), thermoplastic-elastomer tube (Saint-Gobain Co., Ltd., Japan)], pressure transducer (Edwards Lifesciences Co., Ltd., USA), electrical scale (A&D Co., Ltd., Japan), reservoir bottle for the perfusing solution, waste fluid bottle, and digital microscope camera (AnMo Electronics Corp., Taiwan) (Fig. 2 b). The temperature inside the chamber was maintained at 37 °C ± 0.5 °C. The perfusing solutions in the reservoir bottle (4 °C) were infused at the rate of 50 μL/min. The perfusion pressure (arterial pressure) and internal chamber pressure were monitored continuously.
Rodent tail vein catheter
Manufactured by Braintree Scientific
Sourced in Japan, United States
The Rodent Tail Vein Catheter is a specialized medical device designed for intravenous (IV) access in small laboratory animals, such as rodents. It provides a reliable and consistent method for administering fluids, medications, or drawing blood samples from the tail vein of the animal.
Automatically generated - may contain errors
4 protocols using rodent tail vein catheter
1
Ex Vivo Normothermic Kidney Perfusion
2
Intra-Arterial Infusion of Conditioned Media
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Intra-arterial infusion was performed 24 h after MCAO for all experimental groups. Under isoflurane inhalation anesthesia, as described above, the right common carotid artery (CCA), the external carotid artery (ECA) stump and the internal carotid artery (ICA) were exposed, and the pterygopalatine artery was ligated by a 5 ± 0 silk suture. A microcatheter (rodent tail vein catheter with diameter 1F, Braintree Scientific, Inc., Braintree, MA, USA) filled with saline to prevent air bubbles was inserted into the stump of the ECA and advanced into the ICA for 5–6 mm from the bifurcation of the CCA. The catheter external diameter was small enough to allow blood flow around it during the infusion. The catheter was then connected to a 1 mL syringe placed in the microinjector, and 1 mL of concentrated GPC-CM (50 µg/mL of total protein) or NPC-CM (50 µg/mL of total protein), or 1 mL of non-conditioned DMEM/F12-based medium for the control group was delivered into the ICA with the infusion velocity 100 μL/1 min with the maintenance of blood flow in ICA. After intra-arterial administration, the catheter was removed, the ECA stump was electrocoagulated and the incision was closed with a 5 ± 0 silk suture.
3
Anti-Tumor Effects of Erastin and KRIBB3 in SCID Mice
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Indicated HeLa cells were subcutaneously injected into the dorsal flanks right of the midline in SCID mice (weight ~20g). At day seven, mice were injected with Erastin (20 mg/kg/ i.v., twice daily every other day) with or without KRIBB3 (50 mg/kg/ i.p., once daily every other day) for two weeks. The Erastin (C30H31ClN4O4) was bought from Selleck Chemicals. Erastin was dissolved in vehicle (2% DMSO and 98% phosphate buffered saline) and prepared by Ultrasonic Cleaner (Fisher Scientific). A final volume of 300 ul Erastin was applied through the tail vein. The half-life and area under curve (0-t) of Erastin are 27 min and 173 µg.min/ml, respectively. The Rodent Tail Vein Catheter (Braintree Scientific, MTV#1) were used to perform injection. This system allows easier access for both timed injections and repeated bolus administration. Tumors were measured once a week. The volumes were calculated using the following formula: volume (mm3)=length×width2×π/6. This study was approved by the Third Affiliated Hospital of Guangzhou Medical University and the University of Pittsburgh Institutional Animal Care and Use Committees and performed in accordance with the Association for Assessment and Accreditation of Laboratory Animal Care guidelines (http://www.aaalac.org ).
4
Anti-Tumor Effects of Erastin and KRIBB3 in SCID Mice
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