Tumor cells (U87-Luc48 (link)) were implanted into the brain of immunodeficient NMRI-Foxn1nu/Foxn1nu, 8 weeks old male mice (Janvier Laboratories, Laval, France). U87-Luc cells were implanted in the mouse brain by intracerebral injection followed by tumor growth monitoring using bioluminescence (PhotonIMAGER™ systems, BIOSPACE LAB). Mice were anesthetized using intra-peritoneal (i.p.) injection of Xylazine 10 mg/kg and Ketamin 60 mg/kg and then fixed on a stereotactic frame. This framework makes it possible to manipulate the brains of living animals, and to reach isolated areas of the brain precisely relative to markings visible to the naked eye through the use of three-dimensional coordinates. After incising the scalp, the stereotaxic coordinates were calculated for injection of tumor cells into a specific point of the brain, and reproducible for all the mice used. In the study, the tumor cells (5 x 104 cells per mice in 1 μL) were injected at Bregma 0, 2.2 mm to the left of the bregma and 3.2 mm deep to perform the implantation at the level of the striatum.
Nmri foxn1nu foxn1nu
Manufactured by Janvier Labs
Sourced in France
The NMRI-Foxn1nu/Foxn1nu is a laboratory mouse strain. It is a model for studying immunodeficiency and can be used for a variety of research applications.
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7 protocols using nmri foxn1nu foxn1nu
1
Intracranial Implantation of U87-Luc Glioblastoma Cells
2
Xenograft Tumor Growth Inhibition
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To establish treatment cohorts, tumor pieces (1 to 2 mm) from early passage PDXs (≤F4 generation) were soaked in undiluted Matrigel (Becton Dickinson) for 15 to 30 min and subsequently implanted subcutaneously onto female mice (NMRI-Foxn1nu/Foxn1nu, Janvier, St. Berthevin Cedex, France) at two sites (scapular region) using as many as four pieces per site. Tumors were allowed to grow to a size of approximately 100 to 200 mm3, at which time, mice were randomized in the treatment and control groups with five to six mice in each group. Tumor volumes were estimated from 2D tumor measurements by caliper measurements twice a week using the following formula: Tumor volume (mm3) = (π/6)(D)(d2), where d is the minor tumor axis and D is the major tumor axis. Mice were treated daily with JQ1 (Hycultec; 50 mg/kg) by intraperitoneal injection and with irinotecan (Hycultec) intraperitoneally three times per week (Monday, Wednesday, and Friday) at 15 mg/kg, followed by 1 week of treatment pause for two consecutive cycles. OTX015 (Hycultec) was given at 50 mg/kg, i.p., for five consecutive days, followed by 2 days of treatment pause.
3
Xenograft Tumor Growth Inhibition
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To establish treatment cohorts, tumor pieces (1 to 2 mm) from early passage PDXs (≤F4 generation) were soaked in undiluted Matrigel (Becton Dickinson) for 15 to 30 min and subsequently implanted subcutaneously onto female mice (NMRI-Foxn1nu/Foxn1nu, Janvier, St. Berthevin Cedex, France) at two sites (scapular region) using as many as four pieces per site. Tumors were allowed to grow to a size of approximately 100 to 200 mm3, at which time, mice were randomized in the treatment and control groups with five to six mice in each group. Tumor volumes were estimated from 2D tumor measurements by caliper measurements twice a week using the following formula: Tumor volume (mm3) = (π/6)(D)(d2), where d is the minor tumor axis and D is the major tumor axis. Mice were treated daily with JQ1 (Hycultec; 50 mg/kg) by intraperitoneal injection and with irinotecan (Hycultec) intraperitoneally three times per week (Monday, Wednesday, and Friday) at 15 mg/kg, followed by 1 week of treatment pause for two consecutive cycles. OTX015 (Hycultec) was given at 50 mg/kg, i.p., for five consecutive days, followed by 2 days of treatment pause.
4
In vivo Study of NMRI-Foxn1nu/Foxn1nu Mice
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The in vivo study was performed on NMRI-Foxn1nu/Foxn1nu, 8 weeks old male mice purchased from Janvier Laboratories, Laval, France. All animal procedures met the European Community Directive guidelines (Agreement B35-238-40 Biosit Rennes, France/No DIR 13480) and were approved by the local (University of Rennes) ethics committee and ensuring the breeding and the daily monitoring of the animals in the best conditions of wellbeing according to the law and the rule of 3R (Reduce-Refine-Replace).
5
Isolation and Use of Human Pancreatic Islets
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Pancreases not suitable for clinical purposes were obtained with informed written consent from anonymized organ donors [23 (link)]. All the experiments and methods using human pancreatic islets were approved by and performed in accordance with the guidelines and regulations of the regional ethics committee of the Pisa University, Italy and in line with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Human islet isolation and culture was performed as previously described [24 (link)]. The human pancreatic islets or pancreas preparations used in the present study are described in Table S1 .
Six-week old male NMRI-Foxn1nu/Foxn1nu or female SCID CB-17/Icr-Prkdcscid/Rj mice (Janvier Labs, St. Berthevin, France) were used and housed in accordance with the Animal Act 1986/2013, Belgium. The experiments were approved by the Ethical Committee for Animal Welfare (CEBEA) of the ULB, University of Mons, and ULB-Center for Microscopy and Molecular Imaging (CMMI) (Campus Biopole Charleroi, Charleroi, Belgium) (ethical permits CMMI-2013-03;2014, 485N and MU/02/03), Belgium. All applicable institutional and/or national guidelines for the care and use of animals were followed.
Six-week old male NMRI-Foxn1nu/Foxn1nu or female SCID CB-17/Icr-Prkdcscid/Rj mice (Janvier Labs, St. Berthevin, France) were used and housed in accordance with the Animal Act 1986/2013, Belgium. The experiments were approved by the Ethical Committee for Animal Welfare (CEBEA) of the ULB, University of Mons, and ULB-Center for Microscopy and Molecular Imaging (CMMI) (Campus Biopole Charleroi, Charleroi, Belgium) (ethical permits CMMI-2013-03;2014, 485N and MU/02/03), Belgium. All applicable institutional and/or national guidelines for the care and use of animals were followed.
6
Xenografting of Marmoset Testicular Tissue
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In total, 35 nude mice, aged between 5 and 6 weeks were used (NMRI-Foxn1nu / Foxn1nu; Janvier) as recipients for xenografting experiment. The mice were kept at the central animal facility of the medical faculty of the University Münster. Surgical castration of ten nude mice was performed immediately prior to the xenografting procedure. In total, 15 nude mice from intact female (n = 5), intact male (n = 5) and castrated male (n = 5) host groups were used as sham-grafted controls. The remaining 20 nude mice from the three host groups of intact female (n = 5), intact male (n = 10) and castrated male hosts (n = 5) were xenografted with fresh prepubertal marmoset testicular tissue. These mice were kept in the same facility with pellet food and water ad libitum. Mice were anesthetized using ketamine xylazine and six fragments per mouse were ectopically xenografted under the back skin of nude mice using Cancer Implant Needles (GI3 Popper and Sons, Staunton, VA, USA) (4 (link)). All xenografted and sham-grafted mice were kept in groups of 4–5 mice per cage and were provided pellet food and water. Body weight of all mice was recorded weekly before cage change throughout the 20-week grafting period.
7
Mice Models for Tumor and PET Studies
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Healthy female mice weighting 22±2 g (Balbc/Rj, 9 weeks, Janvier, Saint Berthevin, France) were used for ex vivo studies. Immunocompromised female mice weighting 28±2 g (NMRI-Foxn1 nu /Foxn1 nu , 9 weeks, Janvier) were used for tumor growing and in vivo PET studies. The animals were maintained and handled in accordance with the Guidelines for Accommodation and Care of Animals and internal guidelines. All experimental procedures were approved by the Ethical Committee of CIC biomaGUNE and the local authorities (Diputación Foral de Guipúzcoa).
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