Generating PDOX Mouse Model of Colorectal Cancer: Studying Progression of Colorectal Cancer in a Murine Model
Published: April 30, 2023
Abstract
Source: Moret, R. et al. Patient-derived Orthotopic Xenograft Models for Human Urothelial Cell Carcinoma and Colorectal Cancer Tumor Growth and Spontaneous Metastasis. J. Vis. Exp. (2019).
This video describes the protocol for generating a patient-derived orthotopic xenograft colorectal cancer mouse model to study the progression of colorectal cancer cells in non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice. This murine model helps in understanding primary tumor growth and metastases pattern.
Protocol
All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Culturing cell lines Grow HK cells in complete RPMI-1640 medium supplemented with 10% fetal bovine serum, 2 nM glutamine, 100 U/mL penicillin G, and 100 mg/mL streptomycin at 37 °C in a 5% CO2 humidified incubator. NOTE: HK cells are normal human follicular dendritic cells and can …
Divulgaciones
The authors have nothing to disclose.
Materials
| RPMI-1640 Medium | American Type Culture Collection | 110636 | |
| Isoflurane | Henry Schein Animal Health | 108333 | |
| D-Luciferin (150 mg/kg) | Perkin Elmer | 122796 | |
| glutamine (2 nM) | Fisher Scientific | 35050061 | |
| Luc/RFP-lentivirus | From our collaborators. See reference 13: Gills, J. et al. A patient-derived orthotopic xenograft model enabling human high-grade urothelial cell carcinoma of the bladder tumor implantation, growth, angiogenesis, and metastasis. Oncotarget. 9, 32718-32729, doi:10.18632/ oncotarget.26024 (2018). | ||
| penicillin/streptomycin 100 mL (100 U/mL) | Fisher Scientific | 15140-122 | |
| Gas | |||
| 100% Oxygen | Airgas Inc | OX USP200 | |
| 100% CO2 | Airgas Inc | CD USPE | |
| Mice | |||
| 6-8 week old NOD/SCID Mice (female) | Jackson Lab | 1303 | |
| Tools | |||
| 15 mL Conical Tube | Sarstedt | 11799 | |
| 150 mm Tissue Culture Dish | USA Scientific Inc | CC7682-3614 | |
| Forceps | Symmetry Surgical Inc | 06-0011 | |
| Surgical scissors | Symmetry Surgical Inc | 02–2011 | |
| Equipment | |||
| 5% CO2 humidified incubator | Thermo Scientific | 3110 | |
| Bioluminescent (BLI) Imaging Machine | Perkin Elmer | CLS136334 | |
| BLI Imaging Machine Software | Perkin Elmer | CLS136335 | |
| Digital caliper | Fowler Tools and Instruments | 54-115-330 | |
| Dissecting microscope | Precision Instruments LLC | (504) 228-0076 | |
| Isoflurane Induction Chamber | Perkin Elmer | 119038 |
Citar este artículo
Generating PDOX Mouse Model of Colorectal Cancer: Studying Progression of Colorectal Cancer in a Murine Model. J. Vis. Exp. (Pending Publication), e20329, doi: (2023).