Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

The overall goal of this procedure is to observe the development of original and metastatic tumor formation in epithelial ovarian carcinoma.This method can help answer key questions in the gynecologic oncological field, such as understanding the tumor biology or peritoneal defemenation.The main advantage of this technique is that orospatic inoculation of human epithelial ovarian carcinoma, or EOC cells, is performed through a retroperitoneal approach grown with a dorsal flank.To prepare a human ovarian clear cell carcinoma cell line, plate and maintain ES2 cells in RPMI-1640 medium with 10%FBS and PenStrep at 37 degrees Celsius, and 5%CO2.In the logarithmic growth phase, remove the old medium from the culture and use three to four millimeters of PBS to wash the cells once.Next, add one millimeter of trypsin/EDTA and rock the vessel to cover the entire monolayer.Remove and discard the trypsin solution and return the cells to the incubator for three to five minutes.After examining the cells under a microscope to ensure the cells have detached and are floating, add five millimeters of fresh serum-containing growth medium to inactivate the trypsin and re-suspend the cells before transferring them to a 15-millimeter conical tube.Centrifuge the suspension at 300 times G and room temperature