Nude mice

Three previously established and characterized chordoma PDXs were used for in vivo experiments: cluster of differentiation (CD) 3, CD7, and CD39 (35 (link)). The main clinical, histological, and genomic features of our xenografts panel have been previously described (35 (link)). PDX models of chordomas were obtained by engrafting a primary tumor sample into nude mice (Charles River Laboratories). All in vivo experimental procedures, animal care, and housing were performed in accordance with the recommendations of the European Community (2010/63/UE) for the care and use of laboratory animals. Experimental procedures were specifically approved by the ethics committee of the Institut Curie CEEA-IC #118 (Authorization APAFiS# 25870-2020060410487032-v1 given by National Authority) in compliance with the international guidelines.

Female CD1 mice (Charles River, New York, NY) were used for the purification of HF SCs. Female CD1 mice transgenic for krt14-H2B-GFP (Tumbar, 2004 (link)) were used for the purification of TACs. We used Tgfbr2 floxed (Leveen, 2002 (link)) mice to isolate primary keratinocytes. Nude mice were from Charles River Laboratories. For lentiviral injections, transduced mice were confirmed by genotyping with RFP primers: forward 5’ –ATCCTGTCCCCTCAGTTCCAGTAC-3’, reverse 5’-TCCACGATGGT GTAGTCCTCGTTG-3’. For TRE-mycETS2 or mycETS2 (T72D) transduced mice, positive mice were fed with doxycycline-containing chow, starting at P0. Mice were maintained in the Association for Assessment and Accreditation of Laboratory Animal Care-accredited animal facility of The Rockefeller University (RU), and procedures were performed with Institutional Animal Care and Use Committee (IACUC)-approved protocols (#13622-H, #14693-H and #14765-H).

Animal studies were performed as published before [24 (link)] and in accordance to the guidelines of the Canadian Council on Animal Care with institutional certifications (University of British Columbia, A15–0231). Briefly, Caki-1WT/DC cells were injected subcutaneously (5 × 10⁶ cells) in the flank region of 8 weeks old nude mice (Charles Rivers Laboratories, MA, USA). Mice were randomly divided into groups after the tumors reached a volume of 100-200 mm³. Sunitinib malate was suspended in citrate-buffered solution (pH 3.5) and elacridar in diluent (0.5% methyl cellulose and 1% Tween-80 in ddH₂O). Treatment was administered by oral gavage once daily for 5 days followed by 2 days off for 2–3 weeks. For the combination treatment, mice were treated with elacridar 15 min prior to the administration of sunitinib malate. Tumor volume was measured every 3 days using calipers and calculated: tumor volume (mm³) = length×width×height× 0.5. Each treatment group had more than 5 mice. Tumors were fixed with 10% para-formaldehyde (Sigma-Aldrich, MO, USA) for 24-48 h, 70% ethanol for 24 h (VWR International, PA, USA) followed by paraffin embedding.

All animal studies were performed per the guidelines of the IACUC and with protocols approved by the Washington University Division of Comparative Medicine. The TAF15 antibody or isotype control antibody was labeled with IRDye 800CW as per manufacturer’s instructions (Licor). Cancers were induced by injecting A549 or H460 cells in the right hind limbs of nude mice (Charles River). The cancers were irradiated with three fractions of 3Gy or 0Gy (sham) over the course of 24 h. The cancer-bearing mice were then injected with 10 μg of labeled antibodies via the tail vein. Mice were imaged using the Pearl Trilogy small animal imaging system (Licor). Fluorescence was detected using an 800 nm channel. Images were analyzed using the Image Studio software. Background subtracted signal intensity was plotted using Graph Pad Prism software.

Four-week-old male nude mice (Charles River, Beijing, China) were fed under the specific pathogen-free conditions. To generate cell-derived xenografts, mice were randomly divided into groups. A total of 5 × 10⁶ luc-SGC-7901 cells were injected subcutaneously into the mouse right flank subcutaneously. After 7 days, mice were treated with everolimus. The lengths and widths were measured using the vernier caliper at intervals of 7 days. Volume = length × width²/2. After 30 days, 100 μL of 15 mg/mL D-luciferin (Solarbio) was intraperitoneally injected into mice. Tumor load was detected using the In-Vivo Imaging System (PerkinElmer, USA). Mice were sacrificed by CO₂ asphyxia. To investigate the side effects of the therapeutic regimens, we chose weight as the indicator. nude mice without GC transplantation received the same interventions. Mouse weights were measured by an electronic scale at the indicated time. The animal experiments were approved by Animal Center of Chinese PLA General Hospital. Animal experiments conformed to the Animal Research: Reporting of In Vivo Experiments (ARRIVE) guidelines.

All in vivo experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of the Icahn School of Medicine at Mount Sinai. Five-week-old female nude mice (Charles River, MA, USA), weighing 18–20 g and maintained in specific pathogen-free conditions, were used to generate the OC tumor model and to determine the in vivo half-life of the NPs. A total of 1 × 10⁷ OVCAR-3 cells were inoculated subcutaneously into the dorsal region near the hind limb of nude mice (n = 3). When resultant tumors reached approximately 500 mm³, as measured using external calipers, mice were treated with pH-MUC1-Pt-Cy7 NPs via tail vein injection at a concentration of 0.314 mg of Pt (II). A total of 24 h after the injection, in vivo fluorescence imaging was performed as described below. Mice were then euthanized, and subcutaneous tumors and organs (brain, lung, liver, kidney, spleen, and heart) were collected for ex vivo fluorescence imaging and AAS analysis.

For in vivo experiments, 5 × 10⁵ glioma cells were orthotopically transplanted (Stereotaxic coordinate: X(AP) = 1.0 mm, Y(ML) = 2.0 mm, Z(DV) = −3 mm) in Nude mice (Charles River). For bioluminescence imaging, cells were labeled with the lentiviral-based reporter co-expressing RFP and luciferase (SBI). Oral drugs were delivered to xenografted mice once daily by gavage from d2 to d16. Animals were closely monitored for tumor growth, and euthanized when neurologic signs of disease develop. In addition, subsets were sacrificed at appropriate intervals and tissue sections microscopically examined for early evidence of tumor formation. Methods for the analysis of tumor xenograft for morphology, size, proliferation, and differentiation have previously been reported [28 (link),29 (link),30 (link)].