4.3. Ehrlich’s Ascites Carcinoma Cell Line and Induction of In Vivo Solid Tumors

From the Department of Tumor Biology, National Cancer Center, Cairo University, the Ehrlich’s ascites carcinoma (EAC) cell line was obtained. EAC is spontaneous murine breast cancer that acted as the tumor from which an ascites variant was obtained. An ascitic fluid abundant with tumor cells was obtained via intraperitoneal inoculation. Maintenance of the tumor cell line was accomplished in our laboratory by serial intraperitoneal passage into female albino mice at seven- to ten-day intervals. The EAC cells were obtained under strict aseptic conditions. Contamination and viability of EAC cells were evaluated utilizing the Trypan blue dye exclusion technique [64]. EAC cells were suspended in a concentration of 2.5 × 10⁶ EAC cells per 0.1 mL of normal saline. A hemocytometer was used to count the cells under the microscope. Each mouse was inoculated intradermally with 100 μL EAC suspension (2.5 × 10⁶ cells) at the two bilateral sides on the lower ventral region.