Male Lewis rats (LEW/CrlCrlj) were obtained from Charles River Laboratories Japan, Inc. Transgenic rats expressing human heparin‐binding epidermal growth factor‐like growth factor (hHB‐EGF) as a diphtheria toxin receptor under control of the vasopressin promoter were created as described below. Male rats were used for all experiments. Rats were housed under a 12:12 h light/dark photocycle (lights on 7.30 pm) at 22 ± 2°C and 40%–70% relative humidity with food and water available ad lib. Rats were group‐housed before the surgical operation. After surgery, the rats were kept in individual cages. All animal procedures were approved by the Judging Committee of Experimental Animal Ethics of Jichi Medical University and were conducted in accordance with Japanese legislation concerning animal experiments.
Lew crlcrlj
Manufactured by Charles River Laboratories
Sourced in Japan
The LEW/CrlCrlj is a laboratory animal model used in research. It is a specific strain of rat. The core function of this model is to serve as a tool for scientific investigation and analysis, as is typical for laboratory animal models.
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8 protocols using lew crlcrlj
1
Transgenic Rat Model for hHB-EGF Studies
2
Isolation and Culture of Neonatal Rat Cardiomyocytes
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Hearts were removed from LEW/CrlCrlj (Charles River Laboratories Japan, Kanagawa, Japan) or LEW-Tg (Rosa-luc)11Jmsk (Jichi Medical University, Tochigi, Japan) neonatal rats aged 0–3 days old and washed three times with Hanks' Balanced Salt Solution (HBSS; Fujifilm Wako Pure Chemical, Tokyo, Japan) [[30] (link), [31] (link), [32] (link)]. The hearts were minced into pieces less than 2 mm in size, and 7–8 pieces of tissue were placed into a gentleMACS C tube and processed in a gentleMACS dissociator (Miltenyi Biotec, Bergisch Gladbach, Germany). Cardiomyocytes were collected using a Neonatal Heart Dissociation Kit (Milteny Biotec) and selected using a Neonatal Cardiomyocyte Isolation Kit (Milteny Biotec). The number of cardiomyocytes obtained was counted. The cells were seeded on culture dishes coated with fetal bovine serum (FBS; Thermo Fisher Scientific, Tokyo, Japan) and cultured under specific conditions for each experiment.
3
Islet Cell Sheet Transplantation in Diabetic SCID Mice
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All animal experiments were performed with the approval of the Institutional Animal Care and Use Committee of Tokyo Women's Medical University. Male Lewis rats, 8–12 weeks of age (LEW/CrlCrlj, Charles River, Yokohama, Japan) were used as donors for islet cell sheet transplantation. Male severe combined immunodeficient (SCID) mice, 7–10 weeks of age (C.B-17/lcr-scid/scidJcl, CLEA Japan, Tokyo) were used as recipients. Diabetes was induced in the SCID mice by a single intraperitoneal injection of streptozocin (220 mg/kg body weight). Non-fasting blood glucose levels were measured in blood samples collected from the tail vein of each mouse with the use of a handheld blood glucose meter (Glutest Neo Super; Sanwa Chemistry Laboratory, Nagoya, Japan). SCID mice with hyperglycemia greater than a non-fasting blood glucose level of 350 mg/dL were used for the transplantation of islet cell sheets. All animals were kept under a controlled environment (22–24 °C, 55 ± 10% humidity, and a 12-h light/dark cycle).
4
Lewis Rat Experimental Protocol
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A total of 51 female Lewis rats were used in this study. Adult Lewis rats (LEW/CrlCrlj; 8–10-weeks-old, 140–180 g bodyweight-matched) were purchased from Charles River Lab (Yokohama, Japan). They were housed in the Institute for Animal Experimentation, Tohoku University Graduate School of Medicine, under standardized condtions. This study was approved by the ethical committee of the Tohoku University Graduate School of Medicine Committee on Animal Research (No.2015MdA-146).
5
Investigating Vocal Responses in Female Lewis Rats
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Ninety-seven Lewis female rats (LEW/CrlCrlj, Charles River Laboratories Japan, Inc., Kanagawa, Japan) were used. Our previous studies8 (link),9 (link) reported that female Lewis rats emit more USVs than male rats. Thus, we investigated vocal responses of female rats in this study. The rats were obtained from an animal supplier at 3 weeks of age and were housed in pairs under a 12:12 h light/dark cycle (lights on at 7:30 am) at 22 ± 2°C and 55 ± 15% relative humidity. Food and water were available ad libitum. Animal experiments were conducted after receiving approval from the Animal Experiment Committee of Jichi Medical University and were in accordance with the Institutional Regulations for Animal Experiments and Related Activities in Academic Research Institutions under the Jurisdiction of the Ministry of Education, Culture, Sports, Science and Technology.
6
Lewis Rat Neurodevelopmental Study
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Animal experiments were carried out after receiving approval from the Animal Experiment Committee of Jichi Medical University and were in accordance with the Institutional Regulations for Animal Experiments and Fundamental Guidelines for Proper Conduct of Animal Experiments and Related Activities in Academic Research Institutions under the jurisdiction of the Ministry of Education, Culture, Sports, Science and Technology.
Forty-four male rats of the Lewis strain were used in the present study. These animals were produced by mating of pairs of sires and dams obtained from an animal supplier (LEW/ CrlCrlj, Charles River Laboratories Japan, Inc., Kanagawa, Japan) and were weaned at the age of three weeks. After weaning, male rats were housed in pairs and used in the present study. The animals were maintained under a 12: 12 h light/dark cycle (lights on at 7:30 am) at 22 ± 2 °C and 55 ± 15% relative humidity. Food and water were available ad libitum.
Forty-four male rats of the Lewis strain were used in the present study. These animals were produced by mating of pairs of sires and dams obtained from an animal supplier (LEW/ CrlCrlj, Charles River Laboratories Japan, Inc., Kanagawa, Japan) and were weaned at the age of three weeks. After weaning, male rats were housed in pairs and used in the present study. The animals were maintained under a 12: 12 h light/dark cycle (lights on at 7:30 am) at 22 ± 2 °C and 55 ± 15% relative humidity. Food and water were available ad libitum.
7
Female Lewis Rat Breeding Protocol
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Animal experiments were conducted under approval from the Animal Experiment Committee of Jichi Medical University and were in accordance with the Institutional Regulations for Animal Experiments and Fundamental Guidelines for Proper Conduct of Animal Experiments and Related Activities in Academic Research Institutions under the jurisdiction of the Ministry of Education, Culture, Sports, Science and Technology. Animal experiments were performed in accordance with the “Animal Research: Reporting of In Vivo Experiments” (ARRIVE) guidelines (https://www.nc3rs.org.uk/arrive-guidelines ).
Forty-two female Lewis rats were used in this study. These animals were produced in our laboratory by mating of rats obtained from a supplier (LEW/ CrlCrlj, Charles River Laboratories Japan, Inc., Kanagawa, Japan)28 (link). After weaning at the age of three weeks, female rats were housed in pairs at controlled temperature (22 ± 2 °C) and humidity (55 ± 15%) under a 12-h light/dark cycle (lights on at 7:30 am to 7:30 pm). Food and water were available ad libitum.
Forty-two female Lewis rats were used in this study. These animals were produced in our laboratory by mating of rats obtained from a supplier (LEW/ CrlCrlj, Charles River Laboratories Japan, Inc., Kanagawa, Japan)28 (link). After weaning at the age of three weeks, female rats were housed in pairs at controlled temperature (22 ± 2 °C) and humidity (55 ± 15%) under a 12-h light/dark cycle (lights on at 7:30 am to 7:30 pm). Food and water were available ad libitum.
8
Recording Rat Vocalization Responses
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The pleasant call (PC) or distress call (DC) was recorded from an adult female rat (Rattus norvegicus domesticus; LEW/CrlCrlj, Charles River Laboratories Japan). For the recording of PC, the animal was stroked by hand on the experimenter’s lap for around 5 minutes. To elicit DC, a different animal was transferred to a wire-topped experimental cage and habituated to the cage for 5 minutes. Then, the animal received air-puff stimuli (0.3 MPa) with an inter-stimulus interval of 2 s to the nape from a distance of approximately 5–10 cm. Immediately after 30 air-puff stimuli were delivered, USVs were recorded for 5 min. These vocalizations were detected by a microphone placed at a distance of approximately 15–20 cm from the target animal. The detected sound was digitally recorded at a sampling rate of 384 kHz.
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