Human colon cancer HT-29 cells were harvested with the use of 0.05% trypsin/0.02% EDTA, washed twice with serum-free minimum essential medium (α-MEM) and resuspended in Hank’s medium. A single-cell suspension (3.5×106/200 μl per mouse) with cell viability >90% was inoculated subcutaneously (s.c.). For orthotopic transplantation (i.i.), the anesthetized mouse was placed on a wooden board in the right lateral position and the incision was made through the left upper abdominal pararectal line and peritoneum. The cecal wall was carefully exposed, placed and fixed between layers of sterile gauze. A 1–2 mm piece of specimen, derived from a primary tumor grown s.c. in another mouse, was fixed to the serosal part of the cecal wall with 5-0 surgical sutures. After implantation, the peritoneum and abdominal wall were sutured with 4-0 surgical sutures (Dexon-‘S’; Polfa, Poznań, Poland). Tumor cell transplantations were performed under general anesthesia with a mixture of ketamine hydrochloride (100 mg/kg; Ketamina 10%; Biowet, Puławy, Poland) and xylazine hydrochloride (20 mg/kg; XylaRiem; Riemser Arzneimittel AG, Germany).
Ketamina 10
Manufactured by Biowet
Sourced in Poland
Ketamina 10 is a laboratory equipment designed for precise and controlled dosing of ketamine solutions. It features a digital display, adjustable flow rate, and safety mechanisms to ensure accurate and reliable ketamine administration in a laboratory setting.
Automatically generated - may contain errors
6 protocols using ketamina 10
1
Establishment of Human Colon Cancer Xenograft Models
2
Rosmarinic and Sinapic Acid Effects in Rats
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The experiment was carried out on three-month-old female Wistar rats. The experiment was conducted under the approval of the Local Ethics Committee in Katowice (permission numbers: 38/2015, 148/2015, and 66/2016). The rats were purchased at the Center of Experimental Medicine, Medical University of Silesia (Katowice, Poland).
In the course of the experiment the following drugs were administered orally to the rats: rosmarinic acid (Sigma-Aldrich, St. Louis, MO, USA), sinapic acid (Sigma-Aldrich, St. Louis, MO, USA) and estradiol hemihydrate (Estrofem, Novo Nordisk A/S, Bagsvard, Denmark). As anesthetics ketamine (Ketamina 10%, Biowet Puławy, Puławy, Poland) and xylazine (Xylapan, Vetoquinol Biowet, Gorzów Wlkp., Poland) were used.
In the course of the experiment the following drugs were administered orally to the rats: rosmarinic acid (Sigma-Aldrich, St. Louis, MO, USA), sinapic acid (Sigma-Aldrich, St. Louis, MO, USA) and estradiol hemihydrate (Estrofem, Novo Nordisk A/S, Bagsvard, Denmark). As anesthetics ketamine (Ketamina 10%, Biowet Puławy, Puławy, Poland) and xylazine (Xylapan, Vetoquinol Biowet, Gorzów Wlkp., Poland) were used.
3
Wistar Rat Experiment: Sinapic Acid and Estradiol
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The experiment was conducted on mature, 3-month-old female Wistar rats, with the approval of the Local Ethics Commission in Katowice (permission numbers 38/2015, 148/2015, and 66/2016). The animals were provided by the Center of Experimental Medicine, Medical University of Silesia, Katowice, Poland. Rats had unlimited access to drinking water and standard laboratory feed (Labofeed B; Wytwórnia Pasz “Morawski”, Kcynia, Poland). The following substances and drugs were used: sinapic acid (Sigma-Aldrich, St. Louis, MO, USA), estradiol hemihydrate (Estrofem, Novo Nordisk A/S, Bagsvard, Denmark), ketamine (Ketamina 10%, Biowet Puławy, Puławy, Poland), xylazine (Xylapan, Vetoquinol Biowet, Gorzów Wlkp., Poland).
4
Murine Blood Sample Collection
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Male wild-type C57BL/6J mice (weighing 25–27 g) were used in the experiments. These experiments were conducted in accordance with the EU Guidelines on Animal Experiments (European Directive 2010/63/EU), and all procedures involving animals and their care were approved by the Local Ethical Committee on Animal Testing in Olsztyn (Approval No.: 93/2018, date of approval: 18 December 2018). The animals were purchased from the Center for Experimental Medicine in Bialystok. They were anesthetized by intraperitoneally (i.p.) administering a single dose of ketamine (120 mg/kg, Ketamina 10%; Biowet, Poland) and xylazine mixture (12.5 mg/kg, Xylapan; Biowet, Poland). Blood samples were collected from the right heart ventricle of mice with a 3.8% sodium citrate solution (1:10, v/v). After the experiments, the animals were killed by cervical dislocation.
5
Laser-induced thrombosis in diabetic mice
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Male and female C57BL6 mice (weighing 18–23 g) were used in this study. They were randomly divided into 12 groups:
All methods are in accordance with ARRIVE guidelines.
Contr Norm-female—female normoglycemic mice treated with 5% water solution of gum arabic;
Contr STZ (streptozotocin)-female—female diabetic mice treated with 5% water solution of gum arabic;
CANA Norm-female—female normoglycemic mice treated with CANA;
CANA STZ-female—female diabetic mice treated with CANA;
DAPA Norm-female—female normoglycemic mice treated with DAPA;
DAPA STZ-female—female diabetic mice treated with DAPA;
Contr Norm-male—male normoglycemic mice treated with 5% water solution of gum arabic;
Contr STZ-male—male diabetic mice treated with 5% water solution of gum arabic;
CANA Norm-male—male normoglycemic mice treated with CANA;
CANA STZ-male—male diabetic mice treated with CANA;
DAPA Norm-male—male normoglycemic mice treated with DAPA; and
DAPA STZ-male—male diabetic mice treated with DAPA.
All methods are in accordance with ARRIVE guidelines.
6
Dental Implant Osseointegration in Minipigs
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The study was performed on 12 16-month-old minipigs with an average weight of 50-60 kg. The study protocol was approved by the commission for animal studies.
The pigs underwent the removal of the permanent premolar teeth 2 months after the extraction of the deciduous premolar teeth in the jaw. Care was taken to avoid bone wall fractures. The extractions, implantations and euthanasia were performed under general anesthesia by intravenous application of pentobarbital (Vetbutal ® , Biowet Puławy, Poland) at doses of 25 mg/kg along with the premedication ketamine (Ketamina 10%, Biowet Puławy, Poland) and local infiltration of 2% lignocaine with noradrenaline 1:200,000 (Polfa, Poland). Eight weeks after the extractions, a total of 60 implants were inserted in the 12 animals. Each pig was to receive 5 implants (1 per surface state and 2 reference implants) in the mandible (Fig. 1,2). The position of the implants was statistically fully randomized. The implants were placed endosseously under continuous water cooling. After the insertion, the flap was repositioned using resorbable sutures (4/0 Monosyn ® , B. Braun, Melsungen, Germany). The animals were euthanized 12 weeks post-implantation using 50 mg/kg of pentobarbital (Morbital) and block biopsies of the implant sites were collected.
The pigs underwent the removal of the permanent premolar teeth 2 months after the extraction of the deciduous premolar teeth in the jaw. Care was taken to avoid bone wall fractures. The extractions, implantations and euthanasia were performed under general anesthesia by intravenous application of pentobarbital (Vetbutal ® , Biowet Puławy, Poland) at doses of 25 mg/kg along with the premedication ketamine (Ketamina 10%, Biowet Puławy, Poland) and local infiltration of 2% lignocaine with noradrenaline 1:200,000 (Polfa, Poland). Eight weeks after the extractions, a total of 60 implants were inserted in the 12 animals. Each pig was to receive 5 implants (1 per surface state and 2 reference implants) in the mandible (Fig. 1,2). The position of the implants was statistically fully randomized. The implants were placed endosseously under continuous water cooling. After the insertion, the flap was repositioned using resorbable sutures (4/0 Monosyn ® , B. Braun, Melsungen, Germany). The animals were euthanized 12 weeks post-implantation using 50 mg/kg of pentobarbital (Morbital) and block biopsies of the implant sites were collected.
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