Athymic homozygous female nude mice were purchased from Swiss Nu/Nu, Charles River Laboratories, Paris, France and were 5–6 weeks old and weighed between 16 and 18 g. All experimental animals were housed (three per cage) in polycarbonate cages and kept on a 12 h light/dark cycle under specific pathogen-free conditions. Food and water were given ad libitum. Approval for housing and breeding was obtained from the Mansoura Medical Research Ethics Committee of the University of Mansoura (protocol number: RZOO1; approval date: 26 January 2017).
Swiss nu nu
Manufactured by Charles River Laboratories
Sourced in France
The Swiss Nu/Nu is a type of laboratory animal model characterized by the absence of a thymus gland, leading to a lack of mature T cells. This model is commonly used in research applications that require an immunocompromised animal. The core function of the Swiss Nu/Nu is to serve as a tool for studying various diseases and therapeutic interventions in the absence of a functional adaptive immune system.
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11 protocols using swiss nu nu
1
Nude Mice Housing and Handling
2
Ovariectomized Mice for Breast Cancer
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Swiss nu/nu (7 weeks, n = 18) female mice were purchased from Charles River Laboratories-France. Prepubertal mice were ovariectomized under isoflurane anesthesia to remove endogenous ovarian hormone production. Four days before cancer cell injection, mice were implanted s.c. either with a 1.7 mg/60 days MP or with a ME2/60 days RI, or were sham operated (untreated control group). Human adenocarcinoma MCF7 cells were used as previously described [14 (link)]. Tumor cells (1 × 106 cells suspended in 200 μl of Matrigel) were injected s.c. to both flanks of mice. Tumors were measured every 2 to 3 days with digital caliper and tumor volume was calculated as V (mm3) = a x b2× 0.52 (n = 12 tumors per experimental group). At sacrifice, tumors were immediately resected and weighed. The bladder, uterus and kidneys were also dissected, fixed using 4% formalin and embedded in paraffin for histological analysis. Sections were cut at 6 μm, deparaffinised in xylene and rehydrated through graded alcohols, then stained with hematoxylin/eosin. We systematically checked that E2-untreated ovariectomized mice had an atrophied uterus (<10 mg) and that those implanted with an E2-releasing pellet had a significant increase of uterine weight.
3
NOD-SCID-IL2Rγ and Nude Mice Anesthesia
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Three-month-old male NOD-SCID-IL2Rγ (NSG) mice (25–30 g) and 3-month-old nude mice (Swissnu/nu; 25–30 g) from Charles River Laboratories (L'Arbresle, France) were used in this study. Animals were provided with food and water ad libitum and housed in a colony isolator maintained at a constant temperature of 19–22°C and humidity (40–50%) on a 12:12 hr light/dark cycle. The experiments were performed in compliance with the European Communities Council Directive of November 24, 1986 (86/609/EEC) and the principles of laboratory animal care (NIH publication No. 85-23, revised 1985) and were approved by our institutional committee on animal welfare (CETEA-CEA DSV IdF, saisine number #12–029). All surgical procedures were performed under anesthesia with ketamine (75 mg/kg, Imalgen; Merial, Lyon, France) and medetomidine (1 mg/kg, Domitor; Pfizer, Paris, France). After the surgery, paracetamol (1.64 mg/mL, Doliprane; Sanofi, France) was administered in the drinking water for 1 week.
4
Schistosoma mansoni Infection in Nude Mice
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Three athymic homozygous male nude (Foxn1nu−/Foxn1nu−, herein referred to as nude, nu/nu) and six heterozygous female (Foxn1nu−/Foxn1nu+, herein referred to as half-nude or nu/+) mice (Swiss Nu/Nu, Charles River Laboratories, Paris, France) were obtained through the courtesy of Professor Dr. Mohamed Ghoneim, Urology and Nephrology Center, Mansoura, Egypt and were housed (three per cage) in sterilized polycarbonate cages on a 12 h light/dark cycle under aseptic conditions. Food and sterile water were given ad libitum. Approval for housing and breeding was obtained from the Mansoura Medical Research Ethics Committee of the University of Mansoura. Notably, the mice are outbred, not inbred; in addition, the homozygous nude mice lack a thymus, are unable to produce T cells or to mount many types of adaptive immune responses, especially antibody formation, requiring CD4+ helper T cells, and lack hair (nude). The heterozygous mice are immunocompetent and haired (albino).
Cercariae of an Egyptian strain of S. mansoni were obtained from the Schistosome Biological Materials Supply Program, Theodore Bilharz Research Institute (SBSP/TBRI), Giza, Egypt, and used for infection immediately after shedding from Biomphalaria alexandrina snails. Infection of the mice was performed via whole body exposure to viable cercariae as described previously [7] (link), [13] , [16] .
Cercariae of an Egyptian strain of S. mansoni were obtained from the Schistosome Biological Materials Supply Program, Theodore Bilharz Research Institute (SBSP/TBRI), Giza, Egypt, and used for infection immediately after shedding from Biomphalaria alexandrina snails. Infection of the mice was performed via whole body exposure to viable cercariae as described previously [7] (link), [13] , [16] .
5
Xenograft Mouse Model for CiSCs
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All animal procedures were carried out at the Urology and Nephrology Center Animal House in accordance with the institutional and National Institute of Health guidelines for the care and use of laboratory animals. The study protocol was approved by the ethical committee of Mansoura University. Nude mice (Swiss Nu/Nu; Charles River Laboratories, Paris, France) were housed as one mouse per cage. The mice (n = 5 per group) were anesthetized by intraperitoneal injection of ketamine (100 mg/kg) and diazepam (5 mg/kg). A total of 1 × 106 CiSCs were implanted under the kidney capsule. After 2 months, the mice were euthanized and the kidneys were stained for histological analysis.
6
KRAS-Driven Tumor Growth Suppression
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Animal experiments were performed in accordance with the French regulations and approved by the local animal ethics committee (License No. CEEA.2012.84). MCF10A-KRASV12 cells were transduced using either control lentivectors or BCL2L1-targeting sh-RNA (MOI 5). Four days later cells were washed, harvested in PBS and mixed (50:50) in Matrigel (BD Biosciences) before injection in 9-week-old female nude mice (Swiss Nu/Nu, Charles River Laboratories). 7 mice were assayed for each group. We subcutaneously injected into the right flank of mice 5 × 104 cells in a final volume of 150 μl. Importantly, the number of transplanted-KRASV12 cells was determined by serial dilution assays as the minimal to inject in order to obtain 100% tumour uptake in less than 5 months (not shown). The presence of a visible or palpable tumour was then regularly monitored and tumour volume was measured using callipers during a period of 32 weeks. The animals were euthanized when tumour volume reached 2000 mm3 or when signs of tumour necrosis were observed.
7
Xenograft Tumor Induction in Mice
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The study was approved by the Ghent University Ethical Committee on animal experiments (ECD 17/14). All animals (n = 10) were kept and handled according to the European guidelines (Directive 2010/63/EU) and housed under environmentally controlled conditions (12 h normal light/dark cycles, 20–24 °C and 40–70% relative humidity) with food and water ad libitum. On the day of the inoculation, LNCaP and PC-3 cells were washed twice with FBS-free RPMI 1640 medium and two cell suspensions of 5 × 106 cells/100 µL were prepared and kept on ice until inoculation. Four-week-old male athymic nude mice (swiss nu/nu, Charles River Laboratory, France) were subcutaneously injected with 200 µL 1:1 cell:Matrigel suspension using precooled insulin syringes on either side of each mouse (LNCaP, n = 6; PC-3, n = 4) at shoulder height. Tumor growth was monitored weekly for 5–6 weeks until tumors reached a diameter between 5 and 10 mm.
8
Animal Study on Nude Mice
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Animal studies were approved by the Institutional Ethics Committee of the Faculty of Medicine, University of Coimbra (ORBEA_91_2015/08) and were performed according to the local and international guidelines on animal experimentation. Female nude mice (Swiss nu/nu), 6–8 weeks old, were obtained from Charles River Laboratories, (Barcelona, Spain) and housed under pathogen free conditions in individual ventilated cages.
9
Xenograft Tumor Growth in Nude Mice
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ShScr and shNPM1 LNCaP cells were grown to confluency then resuspended in matrigel (BD matrigel basement membrane matrix phenol red free, BD Biosciences, Le Pont de Claix, France) to a concentration of 9×106 cells/ml. Three hundred µl of the cell suspension (approximately 3×106 cells) were injected subcutaneously in 6 weeks old Nude mice (Swiss NU/NU, Charles River, L'Arbresle, France). Disease progression was monitored daily, based on a set of general wellness criteria set by the animal care committee, and body mass was recorded thrice a week until a predetermined endpoint was reached. Endpoints included: dehydration and/or weight loss of over 10%, any evidence of respiratory distress, body weight increase of over 5 g from the average. Tumours were measured thrice a week using an electronic caliper since palpable tumours were detectable. Mice were killed by cervical dislocation under gas anesthesia. Xenografts were then removed, weighted and snap-frozen in liquid nitrogen for RNA and protein analyses.
10
Ex vivo biodistribution of 18F-PSMA-11 in mice
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Eight male athymic nude mice (swiss nu/nu, Charles River Laboratory, France) were subjected to ex vivo biodistribution. One additional mouse bearing LNCaP xenograft was added to evaluate tumor uptake. All mice received 1.95 ± 0.10 MBq 18F-PSMA-11 and were sacrificed at 1 h (n = 4 + 1) or 2 h (n = 4) post injection (p.i.). Excretory organs (kidneys, bladder and liver) and bone fragments (femur, humerus, sternum and skull) were removed, weighted and measured using a gamma counter.
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