All animal studies (primary cell retrieval and in vivo analysis) were approved by the University of Ottawa Animal Care Committee. Sprague-Dawley rats, Fischer rats, and C57BL/6 mice were obtained from Charles River Laboratories (Wilmington, MA, USA).
Fischer rats
Manufactured by Charles River Laboratories
Sourced in United States, France
Fischer rats are inbred laboratory rats commonly used in research. They are a well-established model organism with a consistent genetic background, making them useful for a variety of scientific applications.
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9 protocols using fischer rats
1
Animal Studies on Sprague-Dawley and C57BL/6
2
Liporubicin and Photodynamic Therapy for Sarcoma
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Thirty-eight Fischer rats (Charles River Laboratories, France) underwent subpleural sarcoma implantation in their left lower lobe. This was followed 10 days later by a re-thoracotomy. Tumor L-PDT was performed using Visudyne and laser light. This was directly followed by the administration of Liporubicin, which was allowed to circulate for 1 hour. IFP was measured in tumor and normal lung in 10 and 8 animals, respectively, before and during 1 hour following L-PDT. In a separate set of five animals, TBF was measured in tumors before and during 1 hour following L-PDT. Liporubicin concentration and distribution in tumors and surrounding lung were assessed by epifluorescence microscopy performed on samples embedded in a cryogenic gel (OCT; Electron Microscopy Sciences, Hatfield, PA, USA) in the different treatment groups (n = 5 per group, total = 10). Finally, five animals were used as controls with no L-PDT. In these, all procedures including Visudyne and Liporubicin were injected, but no light was delivered.
3
Ethical Rat Experiment Protocols
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We used adult male Fischer rats (Charles River, Maidstone, UK) weighing 170–250 g for all experiments in this study. Animals were fed a commercial diet and water ad libitum under controlled conditions (22 ± 2 °C, 55% ± 5% humidity, and a 12-h light/12-h dark cycle). All surgical procedures were licensed by the UK Home Office and approved by the University of Birmingham’s Animal Welfare and Ethical Review Board (PPL: 70/08542; date of approval: 12/03/2015). All animal surgeries were carried out in strict accordance with the guidelines of the UK Animals Scientific Procedures Act, 1986, the Revised European Directive 1010/63/EU, and conformed to the guidelines and recommendations of the use of animals by the Federation of the European Laboratory Animal Science Associations (FELASA). Every effort was made to reduce the number of animals employed and to minimize animal discomfort. Pre- and post-operative analgesia was used as standard and with guidance from the named veterinary surgeon.
4
Sepsis Induction in Aged Rat Model
5
Acclimation and Handling of Fischer Rats
6
Epididymal Sperm Analysis in Aging Rats
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Fischer rats (8 weeks of age) were purchased from Charles River (Tokyo, Japan). The rats were kept in a 12/12 h light-dark cycle with free access to standard rat chow (CE-2; Clea, Tokyo, Japan) and tap water. Starting at 10 weeks of age, all the rats had free access to standard laboratory chow (control group) or to an SAC-containing diet (0.45% w/w in the diet), which markedly improved pulmonary fibrosis and type 2 diabetes in our other models.(14 (link),15 (link)) Animals were cared for according to the specifications outlined in the Guiding Principles for the Care and Use of Laboratory Animals–approved by the Authorities of the Local Committee on Experimental Animal Research.
The control animals were killed when they were 15, 25, 50, and 75 weeks of age under anesthesia with urethane (5 g/kg; i.p.). The SAC-treated animals were killed at 75 weeks of age under anesthesia. Blood was collected using heparinized syringes, and the epididymis and testes were dissected. The right epididymis from each rat was used to prepare the sperm solutions. The epididymal fat was also removed and the epididymis was then placed on a paper towel to remove any liquid before being weighed. For the sperm sampling, the cauda epididymis was cut with surgical scissors on the side of the corpus epididymis, where the vas deferens attaches to the epididymis.
The control animals were killed when they were 15, 25, 50, and 75 weeks of age under anesthesia with urethane (5 g/kg; i.p.). The SAC-treated animals were killed at 75 weeks of age under anesthesia. Blood was collected using heparinized syringes, and the epididymis and testes were dissected. The right epididymis from each rat was used to prepare the sperm solutions. The epididymal fat was also removed and the epididymis was then placed on a paper towel to remove any liquid before being weighed. For the sperm sampling, the cauda epididymis was cut with surgical scissors on the side of the corpus epididymis, where the vas deferens attaches to the epididymis.
7
Acclimation of Male Rats
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Isolation and Culture of Rat Mesenchymal Stem Cells
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MSCs were obtained from the femurs and tibias of 344 male Fischer rats (Charles River, France) aged between 7 and 10 weeks. After epiphyses section, the marrow was recovered by centrifugation at 500g for 5 min according to the method described by Peister et al. [14] . The cell suspension taken up in 10 ml of culture medium was filtered through a sieve (70 µm) and then homogenized by successive passages in a 5 ml pipette. Rat MSCs were incubated with α-MEM supplemented with 10% FBS, antibiotics and 50 ng/ml TGFß1 (Sigma-Aldrich). The medium was renewed regularly in order to keep the cells in culture. When the cellular confluence reached 80%, the cells were seeded. After washing using the PBS1 X, the cells were dissociated by the action of 0.25% trypsin -EDTA 1 mM (Invitrogen, France) at 37°C, then, the action of the trypsin was inhibited by the medium culture. After centrifugation for 5 min at 200g, the cells were replaced in culture medium and incubated at 37°C and 5% CO 2 .
9
Glioblastoma Tumor Model in Rats
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Experiments were conducted in accordance with the recommendations of the Canadian Council on Animal Care and the local Ethics Committee. F98 glioblastoma cells were implanted in the right hemisphere of 17 male Fischer rats (254.6 6 15.9 g, Charles River Laboratories) according to a previously published protocol (13) . The animals underwent dynamic PET scans 9-15 d after implantation. All imaging procedures were performed under isoflurane anesthesia with breathing rate and temperature continuously monitored. An automatic injector was used to administer the 18 F-FET solution through a catheter in the caudal vein. Another catheter was inserted either in the caudal artery (n 5 10) or in the femoral artery (n 5 7) and used for manual blood sampling during the PET scan.
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