Table 14.27.2.

Troubleshooting the Lung Orthotopic Model of NSCLC

Problem	Recommended action
Poor tumor take/variable tumor sizes	Ensure cells are >90% viable Maintain cell cultures in logarithmic growth Ensured cells do not exceed 80% confluency Check cells for contamination such as mycoplasma Inject cells within 1 hr of preparing cell suspension Thoroughly agitate cell suspension between loading syringes Optimize cell number Ensure needle is not advanced at too shallow a depth. Needle should be inserted to a depth of approximately 5 mm. Use luciferin solution by recommended expiration date Ensure luciferin solution is properly injected intraperitoneally. Injection should be performed in the right lower quadrant of the abdomen at a 30° angle. The syringe should be aspirated to insure proper placement. Aspiration of blood or other fluid is an indication of improper needle placement.
High surgical mortality	Check dose of anesthesia Keep mice warm post-surgery with heating lamp Check needle placement. Needle should to be inserted to a depth of approximately 5 mm. Pneumothorax, although occurring infrequently, should be evacuated with a 2-ml syringe and a 27-G needle.
Mice develop infection	Use proper aseptic techniques in preparing cell suspension and performing surgical procedure Make sure all surgical instruments are sterilized Thoroughly prepare incision site with betadine and ethanol Check that mouse colony is pathogen free