Tumor Xenograft Assay on Chick CAM

Tumor cells were transplanted to the chorioallantoic membrane (CAM) of fertilized eggs from genetically identical hybrid LB chicks, followed by treatment and humane euthanasia of the chick embryo as described [26 (link), 48 (link)]. After resection, the tumor take rates were calculated from the percent of transplanted tumors that engrafted and the tumor volumes according to the formula Volume = 4/3 × π × r³ (r = 1/2 × square root of diameter 1 × diameter 2) [49 (link)]. Tumor tissue was fixed in Roti Histofix (Carl Roth, Karlsruhe, Germany) for 2-3 days and embedded in paraffin for further analysis.