Nod cg prkdcscid il2rgtm1wjl szj nsg

NSG (NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ) and NSG-SGM3 (NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Tg (CMV-IL3,CSF2,KITLG)1Eav/MloySzJ) mice were purchased from Jackson Laboratories (n = 12 and n = 11). NCG (NOD-Prkdc^em26Cd52Il2rg^em26Cd22/NjuCrl) mice were purchased from Charles River Laboratories (n = 10), and NOG-EXL (NOD.Cg-Prkdc^scid Il2rg^tm1Sug Tg (SV40/HTLV-IL3,CSF2)10-7Jic/JicTac) mice were purchased from Taconic Biosciences (n = 19). All mice were maintained at the Institute of Human Virology at the University of Maryland School of Medicine, Baltimore, USA, and housed in ventilated micro-isolation cages with autoclaved water and irradiated food in a high-barrier facility under specific pathogen-free conditions. For the xenotransplant, human cord blood from an anonymous donor was purchased from LONZA. Only one donor batch was used to reconstitute all four hu-mice strains.
NSG, NSG-SGM3, and NCG mice pups of 4 to 5 days old were irradiated four hours before engraftment with 1.1 cGy for 30 s from an X-ray irradiator. Thereafter, mice were intrahepatically injected with 50 μL of hCD34+ in PBS 1× with 80,000 cells. Since NOG-EXL mice are IL2rgtm1Sug heterozygous and need to be genotyped when they are born, 14–16-week-old mice were conditioned with 25 mg/kg of busulfan given 48 and 24 h intraperitoneally before a vein injection of 50 μL of hCD34⁺ with 80,000 cells.

Care and use of all mice used for this paper was carried out in accordance with UK Home Office regulations, UK Animals (Scientific Procedures) Act of 2013 under UK Home Office licenses which approved this work and is reviewed regularly by the WTSI Animal Welfare and Ethical Review Board and along the ARRIVE guidelines^{65 (link)}. Atm (129S6-Atm^tm1Awb/J; stock no:008671) and NSG (NOD.Cg-Prkdc^scidIl2rg^tm1Wjl/SzJ; stock no: 005557) knockout (KO) mice were imported from Jackson Laboratories. Mouse genotyping was performed from tail biopsies. Mice were maintained in a specific pathogen-free unit on a 12 h light:12 h dark cycle with lights off at 19:30 and no twilight period. The ambient temperature is 21 ± 2 °C, and the humidity is 55 ± 10%. Mice were housed using a stocking density of 3–5 mice per cage (overall dimensions of caging: 365 × 207 × 140 mm³ (length × width × height), floor area 530 cm²) in individually ventilated caging (Tecniplast, Sealsafe 1284L) receiving 60 air changes per hour. In addition to Aspen bedding substrate, standard environmental enrichment of two Nestlets, a cardboard fun tunnel, and three wooden chew blocks are provided. Mice were given water and diet ad libitum.

Bladder cancer xenografts (BL0293 and BL0440) were previously described with corresponding drug sensitivity data [9 (link)]. Frozen samples from Passage 4 were propagated in NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ) immunodeficient mice (The Jackson Laboratory, Bar Harbor, ME, USA) by injecting single cell suspensions (3 × 10⁶ tumor cells administered in 100 µL volume of cell suspension in 1:1 Matrigel (Corning)) subcutaneously (SQ) into the right flank. Tumor burden was assessed by measuring tumor length and width using digital calipers twice per week for up to 10 weeks. Tumor volume was estimated using the formula (length*width²)/2. Tumors were excised when they reached an estimated volume of 1 cm³ or when other IACUC criteria were met. All animal experiments were approved by the Lawrence Livermore National Laboratory Institutional Animal Care and Use Committee and conform to the Guide for the care and use of laboratory animals. Protocol 168 was approved October 14, 2015 by LLNL IACUC to conduct this work.