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).
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).