Tumor Enzymatic Digestion: A Procedure to Generate Tumor Cell Suspension from Patient-derived Xenograft Cancer Mouse Models
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 procedure to obtain tumor cell suspension from patient-derived xenograft or PDX cancer mouse models. Small animal models provide an expeditious, reproducible, and adaptable platform to study tumor progression and different metastatic patterns.
Protocol
All procedures involving animal models 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 an…
Divulgations
The authors have nothing to disclose.
Materials
| Collagenase IV (1.5 mg/mL) | Worthington Biochemical Corporation | LS004189 | |
| Hyaluronidase (20 mg/mL) | Sigma | H3884 | |
| Deoxyribonuclease I (0.1 mg/mL) | Sigma | D4263 | |
| D-Luciferin (150 mg/kg) | Perkin Elmer | 122796 | |
| McCoy’s medium | Life Technologies | 110862 | |
| Hanks Balanced Salt Solution (HBSS) | Fisher Scientific | SH30016.02 | |
| penicillin/streptomycin 100 mL (100 U/mL) | Fisher Scientific | 15140-122 | |
| RPMI-1640 Medium | American Type Culture Collection | 110636 | |
| 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). | ||
| Trypsin/EDTA | Life Technologies | 15400-054 | |
| 40 µm cell strainer | Fisher Scientific | 08-771-1 | |
| 100 µm cell strainer | Fisher Scientific | 08-771-19 | |
| 15 mL Conical Tube | Sarstedt | 11799 | |
| 50 mL Conical tube | Sarstedt | 15762 | |
| 150 mm Tissue Culture Dish | USA Scientific Inc | CC7682-3614 | |
| 96 Well plate | USA Scientific Inc | CC7682-7596 | |
| Forceps | Symmetry Surgical Inc | 06-0011 | |
| Surgical scissors | Symmetry Surgical Inc | 02-0011 | |
| 100% Oxygen | Airgas Inc | OX USP200 | |
| 100% CO2 | Airgas Inc | CD USPE | |
| 6-8 week old NOD/SCID Mice (male) | Jackson Lab | 1303 | |
| 6-8 week old NOD/SCID Mice (female) | Jackson Lab | 1303 | |
| 5% CO2 humidified incubator | Thermo Scientific | 3110 | |
| Centrifuge | Beckman | 366830 | |
| Isoflurane Induction Chamber | Perkin Elmer | 119038 | |
| Serological pipet (10 mL) | Sarstedt | 86.1254.001 | |
| Dissecting microscope | Precision Instruments LLC | (504) 228-0076 | |
| Bioluminescent (BLI) Imaging Machine | Perkin Elmer | CLS136334 | |
| BLI Imaging Machine Software | Perkin Elmer | CLS136334 | |
| Digital caliper | Fowler Tools and Instruments | 54-115-330 | |
| Electrosurgical generator | ValleyLab | FORCE1C20 | |
| Trypan Blue | Sigma | T6146 |
Citer Cet Article
Tumor Enzymatic Digestion: A Procedure to Generate Tumor Cell Suspension from Patient-derived Xenograft Cancer Mouse Models. J. Vis. Exp. (Pending Publication), e20327, doi: (2023).