This video describes a method to engraft lung adenocarcinoma cells in murine lungs by preconditioning the airways with fibrosis-inducing agent–bleomycin. This method significantly enhances orthotopic engraftment of lung cancer cells into murine lungs
Protocol
All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Bleomycin Treatment Plan to inject mice intratracheally with bleomycin 14 days prior to tumor cell engraftment. Thaw bleomycin stock on ice 2 h prior to injection. Once thawed, dilute bleomycin to the desired working concentration (0.02 U/50 µL or 0.005 U/50 µL), and keep it on ice. <stron…
Representative Results
Table 1: List of recommended doses of bleomycin for mouse strains tested. Mouse Strain Cell Line Number of cells injected % engraftment p value Vehicle</stron…
Declarações
The authors have nothing to disclose.
Materials
Rodent Intubation stand
Braintree Scientific
RIS-100
Recommended stand for intratracheal injection
Bleomycin
Sigma
B5507-15UN
CAUTION Health hazard GHS08
Exel Catheter 24G
Fisher
1484121
Remove needle. For intratracheal injection
Ketamine (Ketaset inl 100 mg/mL C3N 10 mL)
Butler Schein
56344
To anesthetize mice
Xylazine
Butler Schein
33198
To anesthetize mice
Syringe 1ml
BD / Fisher
14-823-434
For intraperitoneal injections
Puralube Veterinary Ophthalmic Ointment
BUTLER ANIMAL HEALTH COMPANY LLC
8897
To prevent eye dryness while under anesthesia
MI-150 ILLUMINATOR 150W MI-150
DOLAN-JENNER INDUSTRIES
MI-150 / EEG2823M
To illuminate and visualize trachea
Graefe Forceps, 2.75 (7 cm) long serrat
Roboz
RS-5111
For intratracheal injection
Flask T/C 75cm sq canted neck, blue cap
Fisher / Corning
353135
Phosphate Buffered Saline
Life Technologies
14190-144
0.25% Trypsin-EDTA
Life Technologies
25200-056
Pipet-Lite Pipette, Unv. SL-200XLS+
METTLER-TOLEDO INTERNATIONAL
17014411
Countess Automated Cell Counter
Life Technologies
AMQAX1000
D-Luciferin powder
Perkin Elmer Health Sciences Inc
122799
For luminescent imaging. Reconstitute powder with PBS for a working concentration of 15mg/ mL. Protect from Light