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Cancer Research

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

Published: December 9th, 2016



1Bell Research Center for Reproductive Health and Cancer, 2Department of Obstetrics and Gynecology, Nagoya University Graduate School of Medicine, 3Department of Cancer Biology, Nagoya University Graduate School of Medicine

To clarify the multistep process of peritoneal dissemination of ovarian carcinoma, the current study presents a murine experimental model of original tumor development and peritoneal metastasis via orthotopic inoculation with these tumor cells.

Epithelial ovarian carcinoma (EOC) is associated with a poor prognosis because it shows peritoneal dissemination. To improve the prognosis, it is important to control peritoneal dissemination. However, it is still unclear how tumor cells detach from primary lesions and attach to the mesothelium. The establishment of an appropriate animal model is needed to gain an understanding of the mechanism of peritoneal dissemination in vivo. In the current study, we introduce the process from the local injection of EOC cells into the murine ovarian surface to the development of metastasis, including the peritoneum and distant organs. Female nude mice (BALB/c nu/nu) at 8 weeks of age were used. Under a microscopic field of view, EOC cells (1 x 105 cells/µl of medium-extracellular matrix (ECM)-based hydrogel/unilateral ovary/mouse) were injected into murine ovaries through a retroperitoneal approach from the dorsal flank. This proposed method is a less invasive procedure for the mouse and minimizes damage to the ovary. Here, we describe the methodological steps in the development of the original and metastatic tumor formation of EOC.

Epithelial ovarian carcinoma (EOC) accounts for the highest rate of cancer-related mortality among gynecological malignancies1. EOC is associated with a poor prognosis, primarily due to its late symptomatology, and is often associated with multiple intraperitoneal disseminations and distant metastases2-4. Peritoneal dissemination is a multi-step process. Firstly, tumor cells detach from the primary lesions, and migrate into the abdominal cavity. When the tumor cells attach to the peritoneal mesothelium, they start to invade tissues through the mesothelium5,6. In order to better understand the tumor biology (e.g., cancer progre....

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The treatment protocol follows the guidelines for animal experimentation adopted by Nagoya University.

1. Preparation of Cell Suspensions

  1. Human ovarian clear cell carcinoma cell line.
    1. Maintain ES-2 cells12 in RPMI-1640 medium with 10% fetal bovine serum (FBS) and penicillin/streptomycin (penicillin: 100 units/ml, streptomycin: 0.1 mg/ml). Culture cells at 37 °C in a 5% CO2 humidified incubator.
    2. Collect cells in the logarithmic growt.......

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Six nude mice had their ovaries inoculated with the human clear cell carcinoma cell line ES-2, as described in this protocol. As shown in Figure 1A, after 2 weeks, 5 of the 6 mice had a tumor in the ovary inoculated with ES-2 cells, but there was no tumor in the contralateral ovary without inoculation (used as a control). Moreover, 2 of the 5 mice showed multiple peritoneal disseminations with ascites. Cells metastasize to the liver (Figure 1B). The ovari.......

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Animal models are essential to analyze the mechanisms of tumor development, progression, metastasis, and drug efficacy against cancer cells. Mice bearing human ovarian cancer cells also have been used as an animal model. Here, we describe a less invasive procedure for the administration of ovarian tumor cells at an orthotopic site. Using this procedure, inoculation into the ovary through a retroperitoneal approach from the dorsal flank offers significant advantages over other commonly used techniques. Firstly, orthotopic.......

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The authors thank the members of the Department of Obstetrics and Gynecology and Cancer Biology for their helpful discussions and technical assistance. This study was supported by a Japan Society for the Promotion of Science (JSPS) Grant-in-Aid for Scientific Research (15K15604; to H. Kajiyama).


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Name Company Catalog Number Comments
Matrigel matrix  BD 354234 ECM-based hydrogel
Insulin syringe TERUMO SS-05M2913 1/2cc, 29Gx1/2"
Suturing needle with suture 11mm, 3/8, Nylon6-0
Reflex 7mm Wound Clip Applier CellPoint Scientific 204-1000
 Reflex 7mm wound clips CellPoint Scientific 203-1000

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