Animal studies were approved by the state animal research committee (LANUV, NRW, Germany) and all animals were cared for according to the guidelines set by the Federation of European Laboratory Animal Science Associations. Six- to 8-week-old female NOD.Cg-PrkdcSCIDIl2rgtm1Wjl/SzJ (NOD-SCID gamma; NSG) mice (Charles River Laboratories, Sulzfeld, Germany) were intravenously engrafted with 3.5 × 106 MOLM-14 cells stably expressing a firefly luciferase-EGFP fusion protein (LucEG). Six days later, mice were intravenously injected with 3.5 × 106 N3-, N4-, or CD8-hinged CD19 or CD33 CAR T cells. At days 6, 13, 20, 27, and 34, the persistence of MOLM-14 cells was assessed via luminescence imaging and PB analysis. For luminescence imaging, mice were intraperitoneally injected with D-luciferin (OZ Biosciences SAS, Marseilles, France) and after 5 min luminescence was measured in a Caliper IVIS Lumina II system (PerkinElmer LAS, Rodgau, Germany) with an exposure time of 15 s. PB was drawn from the tail vein, the erythrocytes lysed with BD Pharm Lyse (BD Biosciences), and the samples analyzed on a MACSQuant Analyzer X flow cytometer for EGFP, CD33, and CD45 expression for MOLM-14 cells and BFP, CAR (ΔNGFR), CD3, and CD45 expression for CAR T cells after staining with CD271-PE, CD3-PerCP-Vio700, CD45-APC, and CD33-APC-Vio770 (all from Miltenyi Biotec)
Nod cg prkdcscidil2rgtm1wjl szj nod scid gamma nsg mice
Manufactured by Charles River Laboratories
Sourced in Germany, Japan
The NOD.Cg-PrkdcSCIDIl2rgtm1Wjl/SzJ (NOD-SCID gamma; NSG) mice are an immunodeficient mouse model developed for biomedical research. They possess a severely compromised immune system, lacking functional T cells, B cells, and natural killer cells. This model is commonly used for the study of human tumor xenografts, stem cell transplantation, and other applications requiring a highly immunocompromised host.
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Lab products found in correlation
Pharm lyse, by BD (1 mentions)
Cd45 apc, by Miltenyi Biotec (1 mentions)
Cd271 pe, by Miltenyi Biotec (1 mentions)
Cd3 percp vio700, by Miltenyi Biotec (1 mentions)
Cd33 apc vio770, by Miltenyi Biotec (1 mentions)
D luciferin, by Ozyme (1 mentions)
Caliper ivis lumina 2 system, by PerkinElmer (1 mentions)
2 protocols using nod cg prkdcscidil2rgtm1wjl szj nod scid gamma nsg mice
1
Xenograft Mouse Model for CAR T Cell Therapy
2
Efficacy of NC114 in Colorectal Cancer Xenograft Model
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All experiments involving animals were performed according to protocols approved by the Committee for the Use and Care of Experimental Animals at the Japanese Foundation for Cancer Research (approval No.: 18‐01‐4). Female Nod.Cg‐PrkdcscidIL2rgtm1Wjl/SzJ (NOD scid gamma (NSG)) mice were purchased from Charles River Laboratories Japan (presently The Jackson Laboratory Japan Inc., Kanagawa, Japan) and housed in a specific pathogen‐free animal facility.
After the mice were acclimated to the environment for a week, colorectal cancer SW480 cells (5 × 106) were subcutaneously injected into gender‐ and age‐matched female 6–8‐week‐old mice. When the tumor volume reached 150–200 mm3, the mice were randomly divided into two groups and treated with either NC114 (2 mg per mouse in 200 μL saline containing 8% DMSO and 23% Kollipore HS15) or vehicle by intraperitoneal administration. Assuming a 1‐week clinical dosing schedule, the compound or vehicle was administered a total of nine times as shown in Fig.11A . Tumor size was assessed twice weekly using a caliper, and tumor volumes were estimated as [(long diameter) × (short diameter)2/2]. The mice were weighed daily except on weekends. At the end of the experiment, the mice were sacrificed and tumor tissues were removed and weighed.
After the mice were acclimated to the environment for a week, colorectal cancer SW480 cells (5 × 106) were subcutaneously injected into gender‐ and age‐matched female 6–8‐week‐old mice. When the tumor volume reached 150–200 mm3, the mice were randomly divided into two groups and treated with either NC114 (2 mg per mouse in 200 μL saline containing 8% DMSO and 23% Kollipore HS15) or vehicle by intraperitoneal administration. Assuming a 1‐week clinical dosing schedule, the compound or vehicle was administered a total of nine times as shown in Fig.
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