B16F10 Tumor Growth

• Medium: DMEM, 10% HI-FBS, L-glutamine, optional P/S
• Feed cells the day prior to injection.
• Harvest by trypsinization when dense. Can be confluent. Should not be overgrown ("piling up"). Adjust cell concentration and wash into cold serum-free DMEM (they don't like PBS). On ice until injected.
• "Subdermal" injection probably best, presumably meaning be sure not to inject the underlying muscle, fat, etc.
• Inject into the triangle formed by tenting the skin with fingers. 1 x 10^{5 -} 1 x10⁶ cells in 100 µl serum-free DMEM.
  

Note from Don Senger, 2007_10_22: "Regarding growth state, confluent is OK so long as they haven't been starved.......therefore, they should be fed the day before and not be over confluent and piling up. For injection, probably serum-free DMEM is best; they don't like PBS. And, yes, keep them on ice until injected. Subdermal injection, i.e. not as  deep as s.c. is probably a good bet. I.d. injection is difficult and less reproducible. Sex may matter re: subtle differences but both should be OK. ..."