Method/Protocol for PDX model in ovarian cancer ?

Hello Yang Meng

The problem of using the classical models for cancer drug screening which include cultured human tumor cell lines and rodent xenografts comprising human cells grown subcutaneously in immunodeficient animals, is the artificial nature of tumor cell lines, typically passaged for many generations in enriched culture media. These models generally may not be representative of the genetic and epigenetic heterogeneity of the original primary tumor. I am happy that you have taken up the PDX model for your work which is widely used in oncology drug discovery.

You could transplant tumor samples collected from patients either during diagnostic biopsy or debulking surgery into immunocompromised mice either orthotopically or non-orthotopically.

Orthotopic ovarian cancer mouse model accounts for transplantation of tumor cells either into ovarian bursa (intrabursally, IB), or into mouse peritoneal space (intraperitoneally, IP). The non-orthotopic locations for ovarian cancer include mammary fat pad, sub renal capsule, and subcutaneous (SC) space.

The advantage of using orthotopic model allows one to assess tumor development in a relevant environment and evaluate efficacy in a preclinical tumor model that mimics the disease process in humans. With orthotopic models, we can closely monitor and accurately quantify primary tumor growth, metastatic activity, and response to therapy scenarios. On the other hand, in non-orthotopic models different microenvironment surrounding non-orthotopic tumor grafts can lead to different transplantation outcome or phenotype.

The orthotopic implantation of tumor fragments directly into the organ of origin to better mimic the complexity of human malignancy has become a hot topic as a preclinical model for oncology drug research.

Please find attached a few references that may be helpful for your experiments.

https://pubmed.ncbi.nlm.nih.gov/24398046/

Article A Human Ovarian Carcinoma Murine Xenograft Model Useful for ...

Article Molecular correlates of platinum response in human high-grad...

Good Luck!

Saif Wahid

Patient-derived xenograft (PDX) model for ovarian cancer can be a valuable approach to study drug resistance in a more clinically relevant context. Here is a general outline of the protocol for establishing a PDX model in ovarian cancer:

Patient Tumor Collection: Obtain informed consent from patients undergoing surgery for ovarian cancer. Collect tumor tissue during the surgical procedure. Ensure that the collection is performed in compliance with ethical guidelines and regulations.

Tumor Processing: Process the tumor tissue immediately after collection. Keep the tissue in sterile conditions and transport it to the laboratory on ice in a suitable transport medium, such as Dulbecco's Modified Eagle Medium (DMEM) or RPMI-1640.

Tumor Fragmentation: Cut the tumor tissue into small fragments (1-2 mm³) using sterile scalpels or scissors. Maintain sterile conditions throughout the procedure to avoid contamination.

Mouse Model Selection: Choose immunodeficient mice as recipients for the PDX model. NOD/SCID (non-obese diabetic/severe combined immunodeficiency) mice or NSG (NOD/SCID/IL2Rγnull) mice are commonly used for this purpose due to their lack of functional immune systems.

Mouse Preconditioning: Irradiate the recipient mice (if using NOD/SCID mice) or administer an anti-CD122 antibody (if using NSG mice) to suppress any residual immune activity and create a permissive environment for tumor engraftment.

Tumor Engraftment: Under sterile conditions, subcutaneously or orthotopically implant the tumor fragments into the recipient mice. The orthotopic approach involves implanting the tumor fragments into the ovary or the omentum for ovarian cancer models.

Tumor Monitoring: Monitor tumor growth regularly using calipers or imaging techniques such as ultrasound or bioluminescence imaging. Record tumor size and body weight measurements at appropriate intervals.

Subsequent Passaging: When the tumor reaches a suitable size (typically 500-1000 mm³), excise the tumor from the mouse and repeat the process by fragmenting and implanting it into new recipient mice. This allows for the establishment of subsequent generations of the PDX model.

Experimental Interventions: Utilize the PDX model to study drug resistance by treating the mice with various anticancer agents, alone or in combination. Monitor tumor response, assess drug efficacy, and investigate mechanisms of resistance.

Tissue Preservation: Collect and preserve representative tumor samples from each passage for downstream analyses, such as histopathological examination, molecular profiling, or genomic analysis.

Could you kindly share your email address with me? I have an ebook related to PDX that I would like to forward to you.

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