FulvR-1 and MPF-R cells (1 × 106) were resuspended in 50 ul of extracellular matrix (ECM) from Engelbreth-Holm-Swarm sarcoma (Sigma-Aldrich) and injected orthotopically into the mammary fat pad of 7-week-old female NOG CIEA mice (Taconic) without exogenous estrogen supplements. When the FulvR-1 and MPF-R tumor xenografts reached 100–150 mm3, treatment with CDK4/6i (palbociclib, 50 mg/Kg bodyweight) combined with fulvestrant (100 mg/Kg bodyweight; n = 7 and 9) or vehicle (castor oil and 25% w/v HPB cyclodextrin; n = 8 and 10) was initiated and continued for up to 7 weeks. CDK4/6i was administered by oral gavage once daily for 5 days a week, whereas fulvestrant was administered subcutaneously once a week. At the end of treatment, animals were euthanized, tumors excised, and FFPE. The animal experiment was approved by the Experimental Animal Committee of The Danish Ministry of Justice and was performed at the animal core facility at University of Southern Denmark.
Ciea nog mice
Manufactured by Taconic Biosciences
Sourced in Denmark
The CIEA-NOG mice are a humanized mouse model developed by Taconic Biosciences. These mice have a severely impaired immune system, allowing for the engraftment of human cells and tissues. The CIEA-NOG mice provide a platform for the study of human-specific diseases and the evaluation of novel therapeutics.
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7 protocols using ciea nog mice
1
Orthotopic Breast Tumor Xenograft Model
2
Subcutaneous Tumor Xenograft Assay
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H226 cells stably transduced with DDX56 shRNAs or control shRNAs were harvested, washed in phosphate buffered saline (PBS) and resuspended in extracellular matrix from the Engelbreth-Holm-Swarm sarcoma (Sigma-Aldrich). Tumor cells (1 × 106) were subsequently injected subcutaneously in the right flank of eight-week-old female NOG CIEA mice (Taconic). At study endpoint (week 5), the mice were euthanized by cervical dislocation and tumors were formalin-fixed and paraffin-embedded. All animal experiments were performed at the Animal Core Facility at University of Southern Denmark and approved by The Experimental Animal Committee, The Danish Ministry of Justice.
3
NOG Mice Xenograft Protocol
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All animal experiments were carried out according to the protocols approved by the ethics committee at the Cantonal Veterinary Office of Geneva, Switzerland (authorization N° 1048/3884/3). Female immunodeficient CIEA-NOG mice (10 to 16 weeks old) were purchased from Taconic (Ejby, Denmark) and were used in all in vivo experiments. Mice were maintained in the animal facility of the Geneva University Medical Center, Geneva, Switzerland and used in compliance with the local rules of animal experimentation approved by the Swiss Veterinary Office.
4
Generating Xenograft Interactome from PDX Models
5
Xenograft Model for Biparatopic PSMA×CD3
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Example 8
5-6 week old male immune-deficient CIEA-NOG mice (Taconic) were implanted with 10 million 22Rv1 cells subcutaneously into their lower right flanks, followed by addition of 10 million human PBMCs via tail vein injection one day following tumor implantation. The animals received treatment with 100 μg of multi-specific antibody or vehicle by tail vein injection starting one day after tumor implantation on days 1, 5, 9 and 13. Tumor volume was quantified using calipers and was recorded for 25 days.
6
Xenograft Model for Biparatopic PSMA×CD3
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Example 8
5-6 week old male immune-deficient CIEA-NOG mice (Taconic) were implanted with 10 million 22Rv1 cells subcutaneously into their lower right flanks, followed by addition of 10 million human PBMCs via tail vein injection one day following tumor implantation. The animals received treatment with 100 μg of multi-specific antibody or vehicle by tail vein injection starting one day after tumor implantation on days 1, 5, 9 and 13. Tumor volume was quantified using calipers and was recorded for 25 days.
7
Efficacy of Combination Therapy in Murine Xenograft Model
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Four to 6 week old female CIEA NOG mice were obtained from Taconic Biosciences (Denmark). All mice were housed in the pharmacology department at the Roche Innovation Center Munich (Penzberg, Germany) in compliance with national and international regulations. 5 × 106 MDA-MB231 cells were s.c. injected into the right flank per mouse. When tumors reached ~70 mm3, mice were assigned to treatment and control groups by randomized allocation. Tumor volume was regularly monitored by means of blinded caliper measurement. αCDCP1-EBV_1 (20 mg/kg/week) and αPD1 Ab (5 mg/kg/week) were administered intraperitoneally every third day, starting on day 21. On day 22, 5 × 106 pEBV_1-specific CD8+ T-cells, in vitro expanded from human PBMCs, were injected intravenously into the tail vein together with 1.5 μg IL-15 and 7 μg sIL15Ra-Fc (R&D Systems).
For post-mortem flow cytometric analyses, tumors were harvested and cut into small pieces by means of a scalpel and digested for 15 min at 37°C in RPMI1640 Medium containing 1 mg/mL Dispase II (Roche), 1 mg/mL Collagenase IV (Sigma Aldrich) and 0.1 mg/mL DNase I (Roche). The digest was strained through a 70 μm nylon mash, washed, and subsequently subjected to staining for flow cytometry.
For post-mortem flow cytometric analyses, tumors were harvested and cut into small pieces by means of a scalpel and digested for 15 min at 37°C in RPMI1640 Medium containing 1 mg/mL Dispase II (Roche), 1 mg/mL Collagenase IV (Sigma Aldrich) and 0.1 mg/mL DNase I (Roche). The digest was strained through a 70 μm nylon mash, washed, and subsequently subjected to staining for flow cytometry.
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