The original outbred ddY mouse spontaneously develops IgAN with a variable age of onset. We previously established "grouped ddY" (gddY) mice by the selective mating of ddY mice with the early-onset phenotype for > 20 generations [23 ] (link). All gddY mice were females and maintained in-house. BALB/c mice (Sankyo Labo Service Corporation Inc., Tokyo, Japan), were used as a healthy control comparison for renal pathology and gene expression studies. All the mice were maintained at the animal facility of Juntendo University (Tokyo, Japan) on normal diet (Oriental Yeast Co., Ltd, Tokyo, Japan) and water ad libitum in a specific pathogen-free room. Care was taken in accordance with the NIH Guide for the Care and Use of Laboratory Animals. The experimental protocol of this study was approved by the Ethics Review Committee for animal experimentation of the Juntendo University Faculty of Medicine (permit no. 2 021 268).
Balb c mice
Manufactured by Sankyo Labo Service
Sourced in Japan
BALB/c mice are an inbred strain of laboratory mice. They are commonly used in immunological and cancer research due to their susceptibility to certain diseases and tumors.
Automatically generated - may contain errors
Lab products found in correlation
Nightowl lb981 nc100 system, by Berthold Technologies (2 mentions)
Eclipse ts100 f, by Nikon (2 mentions)
Midazolam, by Fujifilm (1 mentions)
Medetomidine hydrochloride, by Fujifilm (1 mentions)
Penicillin, by Fujifilm (1 mentions)
Streptomycin, by Fujifilm (1 mentions)
Dimethyl sulfoxide (dmso), by Fujifilm (1 mentions)
Sodium hydroxide (naoh), by Fujifilm (1 mentions)
Methanol meoh, by Kanto Chemical (1 mentions)
Dulbecco s modified eagle s medium, by Fujifilm (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Balb 3t3 clone a31, by American Type Culture Collection (1 mentions)
C57bl 6, by Sankyo Labo Service (1 mentions)
In vivo imaging system, by Berthold Technologies (1 mentions)
Indigo2, by Berthold Technologies (1 mentions)
14 protocols using balb c mice
1
Spontaneous IgA Nephropathy in Grouped ddY Mice
2
Hydrogel Synthesis and Cell Culture
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PRX-VBn consisting of α-CDs and PEG (Mn = 5000) capped with 4-vinylbenzyl groups was synthesized as previously described by Arisaka et al. [13 (link)] (Figures S2 and S3 in Supporting Information ). Iodomethane (MeI) was purchased from Tokyo Chemical Industry (Tokyo, Japan). Methanol (MeOH) was purchased from Kanto Chemical (Tokyo, Japan). Collagen sponges for 35 mm culture dishes were purchased from Koken (Tokyo, Japan). Fetal bovine serum (FBS) was purchased from Gibco BRL, Life Technologies (Carlsbad, CA, USA). Ammonium persulfate (APS), butorphanol tartrate, dimethyl sulfoxide (DMSO), Dulbecco’s modified Eagle’s medium (high glucose, with L-glutamine, phenol red, and sodium pyruvate) (D-MEM), 100 U/mL penicillin, 100 µg/mL streptomycin, medetomidine hydrochloride, midazolam, phosphate buffered saline (PBS), sodium hydroxide (NaOH) and N,N,N’,N’-tetramethylene diamine (TEMED) were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). The BALB/3T3 clone A31, a fibroblast cell line established from mouse embryos, was obtained from the American Type Culture Collection (Manassas, VA, USA). BALB/c mice were obtained from Sankyo Labo Service (Tokyo, Japan).
3
Comparative Tissue Fixation Methods
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Fifteen 10-week-old female BALB/c mice (Sankyo Labo Service, Tokyo, Japan) were used in this study. Mice were divided into three groups: five for ALB perfusion fixation, five for 10% neutral buffered formalin perfusion fixation, and five for ALB and 10% neutral buffered formalin immersion fixation. This study was performed according to the regulations of the animal experiment committee of the Showa University (Animal experiment approval number # 09100).
4
Erythrocyte Agglutination Assay for siRNA Lipoplexes
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Blood (0.3 ml) was collected from the jugular vein of one female BALB/c mice (8 weeks of age; Sankyo Labo Service Corp. Inc.) while under anesthesia by an intraperitoneal injection of 50 mg/kg body weight of pentobarbital (Nembutal, Dainippon Pharmaceutical Co., Ltd.). Erythrocytes were collected from the whole blood at 4°C by centrifugation at 300 g for 3 min and resuspended in PBS as a 2% (v/v) suspension of erythrocytes. Non-PEGylated and PEGylated siRNA lipoplexes with 2 µg siRNA were added to 100 µl of 2% (v/v) erythrocyte suspension, respectively. After incubation for 15 min at 37°C, the sample was placed on a glass plate and agglutination was observed by microscopy.
5
Murine Osteomyelitis Model with P. acnes
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Eighteen adult (12-week-old) male BALB/c mice weighing 20–25 g (Sankyo Labo Service, Shizuoka, Japan) were used. The mice were housed five per cage under specific pathogen-free conditions and maintained on a 12-h light/dark cycle with access to food and water. The mice were fed alfalfa-free food (PicoLab® Select Rodent Diet 50 IF/6F, LabDiet, St. Louis, MO, USA). The mice were anesthetized with an intraperitoneal injection of 50 mg/kg of pentobarbital, and the skin on their left knee was sterilized with povidone iodine as previously described20 (link). A skin incision was made over the left knee, and the distal femur was exposed. A drill and 23G needle were used to make a hole at the distal end of the femur20 (link), and a 0.5 × 8-mm titanium alloy bar was inserted into the mouse femur along with an inoculation of P. acnes (1 × 106 CFU in 1 μl medium) (implant group, N = 12)46 (link). The same technique was used for the control group but without the titanium implant (N = 6). The mice were tracked for 6 months. All of the experiments were approved by the Animal Care and Use Committee of Keio University. And, these experiments were carried out in accordance with the approved guidelines.
6
BALB/c Mice Welfare Protocols
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Eight-week-old female BALB/c mice were purchased from Sankyo Labo Service Co., Inc. (Tokyo, Japan) and maintained under pathogen-free conditions. The protocols for the experiments involving animals were approved by the Institutional Animal Experimentation Committee of the Tokyo University of Science (the approval number for animal experiments: Y21022). All experiments involving animals were conducted following the principles and procedures outlined in the National Institutes of Health Guide for the Care and Use of Laboratory Animals and ARRIVE guidelines.
7
Biodistribution of Cy5-siRNA Lipoplexes in Mice
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Ethical approval was obtained from the Institutional Animal Care and Use Committee of Hoshi University (approval no. P22-016). A total of six female BALB/c mice (weight, 18–20 g; age, 8 weeks; Sankyo Labo Service Corporation, Inc.) were housed at 24°C and 55% humidity under a 12/12-h light/dark cycle (lights on at 8:00 a.m.) with ad libitum access to food and water. siRNA lipoplexes with 20 µg of Cy5-siRNA were systemically administered to mice via the lateral tail vein (n=1/siRNA lipoplex). At 1 h after injecting the siRNA lipoplexes, the mice were euthanized by cervical dislocation, and death was confirmed by cessation of the heartbeat. Tissues (lungs, heart, liver, spleen and kidneys) were analyzed through Cy5 fluorescence imaging using the NightOWL LB981 NC100 system (Berthold Technologies GmbH & Co. KG) as previously described (9 (link),13 (link)). Thereafter, the tissue samples were frozen on dry ice and sliced into 16-µm sections. The localization of the Cy5-siRNA was examined using a fluorescence microscope (Eclipse TS100-F; Nikon Corporation).
8
BALB/c Mouse Model for Streptococcus mutans Study
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Thirty-five twelve-week-old male BALB/c mice (Sankyo Labo Service Corporation, INC., Tokyo, Japan) were used. All animals were given a week to adapt to a 12-hour dark/light cycle under constant temperature (21 ± 1°C) with ad libitum access to food and water. All mice cages used were sterilized with the sterilized filtered top, and all procedures for feeding and cleaning mice were performed inside a biological safety cabinet. Experimental procedures were approved by the Animal Care and Use Committee of Tokyo Medical and Dental University (Tokyo, Japan, authorization numbers: A2019-216C2). After one week adaptation period, the mice were inoculated with 0.3 ml of above S. mutans suspension intravenously into the tail vein of mice sedated with medetomidine hydrochloride (0.75 mg/kg; Domitor®, Zenoaq, Fukushima, Japan) subcutaneously injected into the back at a dose of 0.1 ml/10 g mice. The experimental protocol was schematically shown in Figure 1 Figure 1A Figure 1B
9
Pathogen-Free Murine Experiments
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All animals were kept under specific pathogen-free conditions. C57BL/6 and BALB/c mice were purchased from Sankyo Labo Service Corporation. All animal experiments were performed with the approval of the Animal Research Committee of Hokkaido University.
10
Biodistribution of Cy5-siRNA Lipoplexes
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Ethical approval for this study was obtained from the Institutional Animal Care and Use Committee of Hoshi University (approval no. P21-039). A total of 8 female BALB/c mice (weight, 18–20 g; age, 8 weeks; Sankyo Labo Service Corporation, Inc.) were housed at 24°C and 55% humidity under 12/12-h light/dark cycle (lights on at 8:00 a.m.) with food and water ad libitum. siRNA lipoplexes with 20 µg Cy5-siRNA were administered intravenously to mice via the lateral tail vein (n=1/siRNA lipoplex). At 1 h post-injection of siRNA lipoplexes, mice were sacrificed via cervical dislocation; death was confirmed by cessation of heartbeat. Tissue (lung, heart, liver, spleen, and kidney) was analyzed by Cy5 fluorescence imaging using NightOWL LB981 NC100 system (Berthold Technologies GmbH & Co. KG), as previously described (21 (link)). The images were analyzed using IndiGo2 software (version 2.0.1.0) provided with the in vivo imaging system (Berthold Technologies). Following fluorescence imaging, tissue samples were frozen on dry ice and sliced into 16 µm sections. The localization of Cy5-siRNA was examined using a fluorescent microscope (Eclipse TS100-F; Nikon Corporation) with optical filter Cy5 HQ (excitation, 620/60 nm; dichroic mirror, 660 nm; emission, 700/75 nm; Nikon Corporation).
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