Name: Clinically Relevant Transplantable Tumor Models for Studying Metastatic Ovarian Cancer and Therapeutic Response in Mouse (Video) | JoVE Journal
Uploaded: 2026-05-19T14:00:00.000+00:00
Duration: 10 min 45 s
Description: Learn to establish intraperitoneal and intrabursal mouse models for studying metastatic ovarian cancer progression and therapeutic response using bioluminescent imaging techniques.

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A.P. has been supported by the Department of Defense (HT9425-23-1-0351 and HT9425-23-1-0232), National Institutes of Health (R03CA282712), funds from the Ovarian Cancer Alliance of Greater Cincinnati (PRIV0201 and PRIV0219), and UMGCCC American Cancer Society Institutional Research Grant &#8211; IRG-18-160-16. A.P was also supported by grants from the University of Maryland, Baltimore, Institute for Clinical &#38; Translational Research (ICTR), which is funded in part by the National Center for Advancing Translational Sciences (NCATS) Clinical Translational Science Award (CTSA), UM1TR004926. This article was also supported by funds through the National Cancer Institute - Cancer Center Support Grant (CCSG) &#8211; P30CA134274. A.O. was supported in part by NIH grant T32 GM158458.&#160;

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Authors have no disclosures to report.

The ability to mimic complex interactions between tumor cells, microenvironment, and systemic factors makes mouse models an excellent resource to study ovarian cancer progression12. In addition to advancing our understanding of factors that drive ovarian cancer progression and metastasis, in vivo mouse models also play an important role in pre-clinical evaluation of small molecules and treatment strategies for metastatic ovarian cancer12,15</sup...

Despite significant progress in our understanding of factors that drive ovarian cancer progression, ovarian cancer remains the second most lethal gynecologic malignancy in the United States1. Due to a lack of obvious symptoms and the absence of reliable early diagnostic markers, over 70 percent of ovarian cancer patients have advanced metastatic disease at diagnosis2,3. Current therapeutics are not effective in treating metastatic ovarian cancer and consequently, the 5-year survival rates for patients with advanced-stage disease remain less than 30 percent4. A major cause for the lack of therapeutic success is the emergence of chemoresistance5. Therefore, there is a pressing need to identify factors that drive ovarian cancer metastasis and chemoresistance. Addressing this knowledge gap will be critical for developing more effective therapeutic strategies to target this disease. To achieve these goals, we need in vivo models that accurately mimic the key steps of ovarian cancer progression.
While ovarian cancer cell lines and three-dimensional in vitro models have contributed immensely towards our understanding of factors that impact metastasis and therapeutic response, these models have several limitations6,7,8. For instance, these models do not account for the different non-cancerous cells that are part of the tumor microenvironment, nor capture the complex interactions that happen within the tumor microenvironment6,7,8. Further, it is extremely challenging to study systemic factors and their impact on tumor progression and drug response using in vitro models6,7,8. These limitations of in vitro models highlight the need for in vivo models that accurately mimic the various steps in ovarian cancer metastasis, including complex interactions with other cell types and factors within its microenvironment.
Genetically engineered mouse models (GEMMs) offer an in vivo platform wherein tumors arise spontaneously in a natural microenvironment with an intact immune system9. However, they are time-consuming, challenging, and costly to generate and maintain10. Most GEMMs of ovarian cancer also have a long latency period for tumor development, which makes working with them time-consuming and costly, even after they have been generated10. Transplantable models, such as xenograft and allograft models, offer a good balance between biological relevance and experimental ease, and are often much less expensive to generate than GEMMs11. In addition to having lower latency than GEMMs, the ease with which tumor cells can be genetically modified in vitro makes these models convenient and suitable for studying how different genetic factors impact tumor progression10. Due to these benefits, transplantable tumor models have emerged as valuable tools for both basic and translational research in ovarian cancer.
In this report, we describe the protocol for establishing two different transplantable mouse tumor models that are widely used to study metastatic ovarian cancer: (1) the intraperitoneal model and (2) the orthotopic or intrabursal model. Both models can be established as either xenograft or allograft models. In the intraperitoneal model, cancer cells are injected into the peritoneal cavity of the mice. Introducing ovarian cancer cells into the intraperitoneal space mirrors the stage in ovarian cancer metastasis where metastatic ovarian cancer cells are shed from the primary tumor into the peritoneal cavity. Similar to the metastatic process in humans, these transplanted ovarian cancer cells will need to adapt to the hostile peritoneal microenvironment, attach and invade the mesothelial layer that lines the peritoneal cavity, and form metastatic lesions on intraperitoneal organs and tissues such as the omentum, mesentery, liver, and diaphragm. Depending on the cancer cell line used to generate the intraperitoneal model, metastatic tumors can be accompanied by ascites, as observed in late-stage disease in humans. Thus, the intraperitoneal model has been used widely to understand factors that drive ovarian cancer metastasis and their progression into late-stage disease12. These models are also useful to determine the efficacy of novel therapeutic strategies for metastatic ovarian cancer12. In the intrabursal or orthotopic model, cancer cells are implanted into the bursa, a membranous structure that surrounds the mouse ovary. This model mimics early stages of ovarian tumorigenesis and metastasis. At this stage cancer cells are still confined to the fallopian tube and the ovary. It is from these sites that cancer cells are shed into the peritoneal cavity, ultimately metastasizing to other organs. Therefore, the intrabursal model is useful in studying both early stages of ovarian tumorigenesis as well as events that lead to their subsequent transcoelomic metastasis through the peritoneal cavity12.
Thus, while the both intraperitoneal and intrabursal models are excellent tools to study ovarian cancer metastasis and therapeutic response, they differ in the specific stage along the metastatic process that they mimic. While the intrabursal models can mimic the progression of ovarian cancer from early stages to later stages in the metastastic process, due to the latency associated with this process, they are usually used to study early events in ovarian cancer metastasis. For studies focusing on later stages of ovarian cancer metastasis, the intraperitoneal model is more suitable. Further, the intrabursal model is also technically more challenging to establish compared to the intraperitoneal model12. Establishing the intrabursal model requires access to a surgical suite equipped with microsurgical equipment that will allow precise injection of cancer cells into the ovarian bursa. The ability of cancer cells to establish tumors (&#8220;take rate&#8221; or &#8220;engraftment rate&#8221;) is also usually lower in intrabursal models as compared to intraperitoneal models. Moreover, as tumors cannot be easily observed after orthotopic or intraperitoneal injections, a non-invasive bioluminescent imaging technique is typically employed to monitor tumor progression. Therefore, both intraperitoneal and intrabursal models require additional equipment that will allow non-invasive imaging of tumors12. Ultimately, the experimental goals, availability of resources, and technical skill set will determine the choice of model for each study.
This report describes the step-by-step protocol for establishing intraperitoneal and intrabursal mouse xenograft models using human ovarian cancer cell lines. These models can be established by transplanting the cells into immunocompromised mouse strains such as NSG (NOD.Cg-Prkdcscid&#160;Il2rgtm1Wjl/SzJ) or the Foxn1 nude (B6.Cg-Foxn1 nu/J). The same method can be adapted to generate allograft (syngeneic) models. While establishing allograft models, mouse ovarian cancer cells are implanted into immunocompetent mice of the same genetic background. As allograft models are developed using immunocompetent mice, they are useful for studying how different factors regulate the ovarian tumor microenvironment, the role of the immune system in ovarian cancer progression, and the effectiveness of immunotherapy. In addition to establishing the intraperitoneal and intrabursal xenograft mouse models, this report also describes how to use bioluminescent imaging to non-invasively monitor tumor progression in these models.

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clinically relevant transplantable tumor models for studying metastatic ovarian cancer and therapeutic response in mouse

Clinically Relevant Transplantable Tumor Models for Studying Metastatic Ovarian Cancer and Therapeutic Response in Mouse

Metastatic ovarian cancer is an extremely lethal gynecologic malignancy that lacks effective treatment options. Development of clinically translatable therapeutic approaches for ovarian cancer patients is contingent on understanding factors that drive ovarian cancer metastasis. While most solid tumors exhibit hematogenous metastasis, ovarian cancer cells spread primarily through the intraperitoneal route. This process involves detachment and shedding of cancer cells from the primary tumor into the peritoneal cavity, which then acquire properties that enable them to resist anoikis and form multicellular aggregates. These cell aggregates then attach to and invade the mesothelial lining of the peritoneum, forming metastatic lesions. Clinically relevant in vivo models are essential for understanding how cellular factors and the tumor microenvironment impact the different steps in the metastatic process and for testing novel therapeutic strategies. Two transplantable mouse tumor models widely used to study ovarian cancer metastasis and therapeutic response are the intraperitoneal and intrabursal models. In this report, we describe the step-by-step methodology for establishing these models and highlight their advantages and limitations. Additionally, we describe how to use bioluminescent imaging to non-invasively monitor tumor progression using these models.

Representative data from an intraperitoneal ovarian cancer model using luciferase-tagged OVCAR8 cells in immunocompromised Foxn1 nude mice is shown in Figure 4. Stable luciferase expression in these cells enabled longitudinal monitoring of tumor progression by non-invasive bioluminescent imaging using the IVIS imager as described in section III. Intraperitoneal injection of 5 million OVCAR8-luciferase cells into Foxn1 nude mice resulted in the establishment of metastatic tumors in the perito...

Here we present a step-by-step protocol for establishing the intraperitoneal and the intrabursal tumor models to study ovarian cancer cancer progression, metasatsis, and therapeutic response. Further, we describe how to use luciferase-based non-invasive in vivo imaging in these models to monitor tumor progression in mouse.

Metastatic ovarian cancer is an extremely lethal gynecologic malignancy that lacks effective treatment options. Development of clinically translatable ...

Our research focuses on identifying the molecular factors that drive ovarian cancer progression and therapeutic resistance. These models can be used to investigate how various factors impact ovarian cancer progression and also test new therapeutic strategies.To begin, obtain ovarian cancer cell culture. Once the cells reach 80% to 85% confluency aspirate the medium from the tissue culture plate. Wash the adherent cells twice with sterile 1x PBS without calcium and magnesium. Aspirate the PBS from the tissue culture plate. Add 0.05% trypsin-EDTA to the tissue culture plate. Transfer the plate with cells to the 37 degrees Celsius water-jacketed carbon dioxide incubator used for tissue culture, and incubate for one to five minutes. Once the cells are detached, add FBS-containing media to the tissue culture plate at a ratio of 6 milliliters of media for every 1 milliliter of trypsin used. Gently pipette up and down to dislodge the cells from the tissue culture plate and dissociate them into a single cell suspension. Collect the cells in a conical tube and centrifuge at 400 g for five minutes at room temperature. After removing the trypsin-containing supernatant, resuspend the cell pell

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Learn to establish intraperitoneal and intrabursal mouse models for studying metastatic ovarian cancer progression and therapeutic response using bioluminescent imaging techniques.

Clinically Relevant Transplantable Tumor Models for Studying Metastatic Ovarian Cancer and Therapeutic Response in Mouse

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