Tumor Development on Chick Chorioallantoic Membrane

Tumor development on CAM was performed as previously described [29 (link)]. Briefly, fertilized eggs (animal facility of the University of Geneva, Geneva, Switzerland) were incubated at 38 °C with 80% relative humidity and periodic rotation. On egg development day (EDD) 4, the eggs were drilled at their narrow apex, and the hole was closed with adhesive tape. The eggs were then incubated at 38 °C with 80% relative humidity without rotation. On EDD8 the hole in the eggshell was enlarged to allow access to the CAM. After gently scratching the membrane with a needle tip, SKOV3-Red, SKOV3-Green, or SKOV3-M cells suspension (2 × 10⁶ cells in 30 μL of geltrex, Thermo Scientific) was inoculated, and the hole was hermetically covered with Parafilm®. Eggs were returned to the incubator to allow tumor growth. Tumor growth was then monitored at EDD10.5 and 13.5 using a Wild Heerbrugg M3Z microscope at 10x magnification with a Lumenera INFINITY2-1 CDD camera with Infinity Capture Software.