All animal procedures were approved by the Animal Welfare and Ethical Review Body at Queen Mary University of London and by the UK Home Office in accordance with EU Directive 2010/63/EU. Female Fox Chase SCID mice (Charles River, UK) were 6–8 weeks old on arrival. A first batch of 8 mice was shaved on the lower back (1 cm2), local anaesthetic cream (lidocaine and prilocaine) applied, and a≈10 μL fragment of primary human colon adenocarcinoma tissue (from patient CR-IGR-034P, tumour collection CReMEC, Oncodesign, [22 (link)]) injected subcutaneously in an intrascapular position by trocar under isoflurane anaesthesia. The wound was closed with spray plaster, and animals were monitored more frequently for 2 days. Once tumours had grown to 8–10 mm diameter, one animal was sacrificed each week, the tumour excised and cut into ≈10 μL pieces for direct passaging into a batch of 20–24 animals (5 batches in total), following the same inoculation procedure as above. All animals underwent regular body weight and tumour size measurements and examinations for any signs of abnormalities.
Fox chase scid mice
Manufactured by Charles River Laboratories
Sourced in United States
The Fox Chase SCID mice are a type of genetically modified laboratory mouse model. They are characterized by a severe combined immunodeficiency (SCID) phenotype, which results in a lack of functional T cells and B cells. This model is commonly used in research applications that require an immune-deficient environment.
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Lab products found in correlation
C57bl 6j, by Charles River Laboratories (2 mentions)
C57bl 6j sting1 j sting golden ticket mouse, by Jackson ImmunoResearch (2 mentions)
Balb c mice, by Charles River Laboratories (2 mentions)
Aprotinin, by Bayer (1 mentions)
Qiaamp dna mini kit, by Qiagen (1 mentions)
Athymic nu nu mice, by Charles River Laboratories (1 mentions)
Microvette cb300 capillary blood collection tubes, by Sarstedt (1 mentions)
Kolliphor el, by Merck Group (1 mentions)
12 protocols using fox chase scid mice
1
Xenograft Model of Colon Cancer
2
Murine Multiple Myeloma Xenograft Models
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All animal studies were performed in accordance with the Institutional Animal and Use Committee of Washington University School of Medicine. Mice were anesthetized for all treatments and imaging with 2% v/v isoflurane/100% O2. Female 1–3 month old fox chase SCID mice (Charles Rivers Laboratories) were injected with 3 × 106 MM.1S-GFP-luc cells in 100 μL 1X PBS SQ or IV via lateral tail vein. Tumor burden was monitored weekly via BLI prior to administration of DARA conjugates in both mouse models. For imaging studies in MM.1S SQ and IV mice, mice were randomized into respective cohorts when a mean bioluminescence flux of 1 × 109 photons/second was achieved.
3
Orthotopic Glioma Mouse Models for MRS
4
Engineered Skin Graft Generation
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A fibrin-based matrix containing RDEB fibroblasts was prepared as described (Larcher et al, 2007 (link)). A total of 105 keratinocytes were then seeded on top of the fibrin gel, grown to confluence in culture medium supplemented with 150 IU/ml aprotinin (Trasylol, Bayer) and transplanted onto the back of 8- to 10-week-old Fox-Chase SCID mice (Charles River Laboratories) following the protocol detailed in the Supplementary Materials and Methods. Animal work was authorised by the veterinarian canton de Vaud authorization 2033. Animals were handled according to ethical standards by qualified persons. Immunodeficient mice (SCID and athymic) were housed in an official animal facility, in compliance with Swiss governmental guidelines. Studies were monitored using an online organisational tool. Grafts were harvested at different time points and processed for histology, immunocytochemistry or electron microscopy. All experiments performed conform to NIH, MRC and ARRIVAL guidelines for animal welfare.
5
Generating Human Skin Grafts from RDEB Fibroblasts
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A fibrin‐based matrix containing RDEB fibroblasts was prepared as described previously (Larcher et al, 2007 (link)). Next, 105 keratinocytes were seeded on top of the fibrin gel, grown to confluence in culture medium supplemented with 150 IU/ml aprotinin (Trasylol, Bayer) and transplanted onto the back of 8–10‐week‐old Fox‐Chase SCID mice (Charles River Laboratories) as described previously (Droz‐Georget Lathion et al, 2015 (link)). The mice were handled according to the Canton de Vaud veterinarian guidelines (authorization 2033). The grafts were harvested after 10 weeks and processed for histology and immunocytochemistry. To confirm the human origin of the regenerated epidermis, sections were immunostained for human HLA class I (1:500—SM2012P Acris) using standard protocols.
6
Xenograft Tumor Formation and Dissemination
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A total of 106 transduced keratinocytes or SCC-13 cells were inoculated subcutaneously into the ventral flanks of 7- to 9-week-old athymic Swiss Nu−/− mice (Charles Rivers Laboratories) with a 21-gauge needle. Tumour formation was monitored twice a week and their diameter measured. For dissemination experiments, internal organs of immunodeficient Fox-Chase SCID mice (Charles River Laboratories) transplanted with recombinant COLVII keratinocytes were harvested at the termination of the experiment. DNA was extracted using the QIAamp DNA mini kit (Qiagen) according to the manufacturer's instructions. One hundred nanograms of DNA was submitted to PCR (BioConcept) amplification with GoTaq PCR reagent kit (Promega) with primers listed in the Supplementary Materials and Methods. PCR products were sequenced (Fasteris, Switzerland).
7
Animal Infection Protocols for Immunological Research
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Experimental procedures involving live animals were carried out in agreement with the protocols approved by the Institutional Animal Care and Use Committee (IACUC) at The Johns Hopkins University School of Medicine. For animal infection protocols, pathogen-free age 4-6 weeks female C57BL/6J (Charles River Laboratories, North Wilmington, Mass.), C57BL/6J-Sting1/J (STING−/− Golden ticket mouse) (The Jackson Laboratory, ME, US), Fox Chase SCID mice (Charles River Laboratories North Wilmington, Mass.) and BALB/c mice (Charles River Laboratories, North Wilmington, Mass.) were purchased and housed under pathogen-free conditions at an Animal Biosafety Level-3 (ABSL3) or Biosafety Level-2 (ABSL2) animal facility without cross-ventilation. Fischer 344 female rats age 8 weeks (Harlan, avg. weight 160g) were housed at an BSL2 animal facility. Animals were housed under standard housing conditions (68-76°F, 30-70% relative humidity, 12-12 light-dark cycle) with free access to water and standard chow and were monitored daily for general behavior and appearance by veterinary specialists.
8
Xenograft Modeling of Breast Cancer
9
Intracranial Tumor Xenograft Treatment
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All studies were performed under UCSF Institutional Animal Care and Use Committee approval (AN184161-01E). 6–9 week old female athymic nu/nu mice (20–25 g; Charles River Laboratories, Wilmington, MA, USA) were intracranially injected with ~3 × 105 U87IDHmut cells. 6–9 week old female SCID mice (19–21 g; Fox Chase SCID mice, Charles River Laboratories, Wilmington, MA, USA) were intracranially injected with ~1 × 105 of either BT257 or SF10417 cells from serial orthotopic xenografting as described [49 (link)]. Tumor size was evaluated using MRI. Once tumors reached 2–3 mm in diameter, a baseline set of MR studies was performed (see below). This time point was considered day zero (D0). Mice were then randomized into three treatment groups and treated daily (Monday to Friday) per os (p.o.) with one of the following: (1) 50 mg/kg AG-881 [38 (link)]; (2) 150 mg/kg BAY-1436032 [43 (link)]; (3) 4 mL/kg Ora-plus (for controls). MR studies were then repeated at regular intervals depending on the tumor model and continued until the animal had to be sacrificed (Figure 1 ).
10
Xenograft Modeling of Breast Cancer
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Our mouse xenograft experiments were done following our UTSW IACUC approved mouse protocol (Sieber protocol# 2020-102956). A 14-hour light/10-hour dark cycle facility maintained at 22°C and 50%humidity. Mice were fed a standard chow diet. Female Fox Chase SCID mice (Charles River) (3 months of age) were anesthetized and injected with 1 Million MCF7 cells into mammary tissue below the 4th nipple on the left side. Estradiol was administered at (4mg/kg) bodyweight weekly to support tumor growth. Tumor size was measured twice weekly to track tumor growth. In line with our IACUC protocol we did not allow tumors to exceed 2cm in diameter. Once tumors reach roughly 40mm2 tumor volume, they were collected, weighed, and used for subsequent metabolite measurements. These experiments were done with assistance from the UTSW animals resource center and reflect 2 independent sets of injections from 2 independent isolations of recurrent MCF7 cells. A total of 16 mice/condition were assayed.
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