Name: Establishment and Maintenance of Patient-derived Prostate Cancer Organoids: A Detailed Experimental Protocol (Video) | JoVE Journal
Uploaded: 2025-11-14T13:00:00.000+00:00
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Description: Learn to establish patient-derived prostate cancer organoids using optimized culture protocols. This method preserves tumor fidelity for personalized drug screening and precision medicine research.

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The authors gratefully acknowledge the support of the National Natural Science Foundation of China (Grant No. 82172831) and the Key Research Project of the Jiangsu Provincial Health and Family Planning Commission (Grant No. X20240121).

<ol><li>James, N. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+lancet+commission+on+prostate+cancer%3A+planning+for+the+surge+in+cases%5BTitle%5D)+AND+%22Lancet%22%5BJournal%5D)">The lancet commission on prostate cancer: planning for the surge in cases.</a> Lancet. 403 (10437), 1683-1722 (2024).</li>
<li>Knutson, T. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((AR+alterations+inform+circulating+tumor+DNA+detection+in+metastatic+castration+resistant+prostate+cancer+patients%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">AR alterations inform circulating tumor DNA detection in metastatic castration resistant prostate cancer patients.</a> Nat Commun. 15 (1), 10648(2024).</li>
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<li>Zhou, L., Zhang, C., Zhang, Y., Shi, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Application+of+organoid+models+in+prostate+cancer+research%5BTitle%5D)+AND+%22Front+Oncol%22%5BJournal%5D)">Application of organoid models in prostate cancer research.</a> Front Oncol. 11, 736431(2021).</li>
<li>Gao, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Organoid+cultures+derived+from+patients+with+advanced+prostate+cancer%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">Organoid cultures derived from patients with advanced prostate cancer.</a> Cell. 159 (1), 176-187 (2014).</li>
<li>Drost, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Organoid+culture+systems+for+prostate+epithelial+and+cancer+tissue%5BTitle%5D)+AND+%22Nat+Protoc%22%5BJournal%5D)">Organoid culture systems for prostate epithelial and cancer tissue.</a> Nat Protoc. 11 (2), 347-358 (2016).</li>
<li>Puca, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Patient-derived+organoids+to+model+rare+prostate+cancer+phenotypes%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Patient-derived organoids to model rare prostate cancer phenotypes.</a> Nat Commun. 9 (1), 2404(2018).</li>
<li>Pamarthy, S., Sabaawy, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Patient-derived+organoids+in+prostate+cancer%3A+improving+therapeutic+efficacy+in+precision+medicine%5BTitle%5D)+AND+%22Mol+Cancer%22%5BJournal%5D)">Patient-derived organoids in prostate cancer: improving therapeutic efficacy in precision medicine.</a> Mol Cancer. 20 (1), 125(2021).</li>
<li>Karkampouna, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Patient-derived+xenografts+and+organoids+model+therapy+response+in+prostate+cancer%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Patient-derived xenografts and organoids model therapy response in prostate cancer.</a> Nat Commun. 12 (1), 1117(2021).</li>
<li>Driehuis, E., Kretzschmar, K., Clevers, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Establishment+of+patient-derived+cancer+organoids+for+drug-screening+applications%5BTitle%5D)+AND+%22Nat+Protoc%22%5BJournal%5D)">Establishment of patient-derived cancer organoids for drug-screening applications.</a> Nat Protoc. 15 (10), 3380-3409 (2020).</li>
<li>Neal, J. T., Kuo, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Organoids+as+models+for+neoplastic+transformation%5BTitle%5D)+AND+%22Annu+Rev+Pathol%22%5BJournal%5D)">Organoids as models for neoplastic transformation.</a> Annu Rev Pathol. 11, 199-220 (2016).</li>
<li>Waseem, M., Wang, B. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Organoids%3A+an+emerging+precision+medicine+model+for+prostate+cancer+research%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">Organoids: an emerging precision medicine model for prostate cancer research.</a> Int J Mol Sci. 25 (2), (2024).</li>
<li>Moschovas, M. C., Patel, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neurovascular+bundle+preservation+in+robotic-assisted+radical+prostatectomy%3A+how+I+do+it+after+15%2C000+cases%5BTitle%5D)+AND+%22Int+Braz+J+Urol%22%5BJournal%5D)">Neurovascular bundle preservation in robotic-assisted radical prostatectomy: how I do it after 15,000 cases.</a> Int Braz J Urol. 48 (2), 212-219 (2022).</li>
<li>Li, Y., Qin, M., Liu, N., Zhang, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Organoid+development+and+applications+in+gynecological+cancers%3A+the+new+stage+of+tumor+treatment%5BTitle%5D)+AND+%22J+Nanobiotechnology%22%5BJournal%5D)">Organoid development and applications in gynecological cancers: the new stage of tumor treatment.</a> J Nanobiotechnology. 23 (1), 20(2025).</li>
<li>Xiang, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Building+consensus+on+the+application+of+organoid-based+drug+sensitivity+testing+in+cancer+precision+medicine+and+drug+development%5BTitle%5D)+AND+%22Theranostics%22%5BJournal%5D)">Building consensus on the application of organoid-based drug sensitivity testing in cancer precision medicine and drug development.</a> Theranostics. 14 (8), 3300-3316 (2024).</li>
<li>Van Hemelryk, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Patient-derived+xenografts+and+organoids+recapitulate+castration-resistant+prostate+cancer+with+sustained+androgen+receptor+signaling%5BTitle%5D)+AND+%22Cells%22%5BJournal%5D)">Patient-derived xenografts and organoids recapitulate castration-resistant prostate cancer with sustained androgen receptor signaling.</a> Cells. 11 (22), 3632(2022).</li>
<li>Zhao, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Patient-derived+bladder+cancer+organoids+as+a+valuable+tool+for+understanding+tumor+biology+and+developing+personalized+treatment%5BTitle%5D)+AND+%22Adv+Sci+%28Weinh%29%22%5BJournal%5D)">Patient-derived bladder cancer organoids as a valuable tool for understanding tumor biology and developing personalized treatment.</a> Adv Sci (Weinh). 12 (13), e2414558(2025).</li>
<li>Polak, R., Zhang, E. T., Kuo, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cancer+organoids+2.0%3A+modelling+the+complexity+of+the+tumour+immune+microenvironment%5BTitle%5D)+AND+%22Nat+Rev+Cancer%22%5BJournal%5D)">Cancer organoids 2.0: modelling the complexity of the tumour immune microenvironment.</a> Nat Rev Cancer. 24 (8), 523-539 (2024).</li>
</ol>

The authors have nothing to disclose.

The establishment of PDOs for PCa represents a critical advancement in preclinical modeling, offering significant advantages over conventional 2D cell cultures and PDXs16. The method presented in this study enables the generation of organoids that faithfully recapitulate the histological, genetic, and phenotypic characteristics of the original tumor. By preserving tumor heterogeneity, including AR signaling and cellular architecture, the organoids provide a more biologically relevant platform for ...

Prostate cancer (PCa) remains one of the most prevalent malignancies in men worldwide, second only to lung cancer in cancer-related mortality. According to recent epidemiological studies, the global incidence of PCa has been steadily rising, and it is projected that the number of new cases of prostate cancer annually will rise from 1.4 million in 2020 to 2.9 million by 20401. Despite advancements in early detection and the development of novel therapies, including androgen deprivation therapy (ADT) and targeted agents, prostate cancer remains a significant clinical challenge due to its heterogeneous nature and the inevitable development of treatment resistance, particularly in advanced stages2. Castration-resistant prostate cancer (CRPC), which emerges after ADT failure, represents a lethal form of the disease, characterized by poor survival outcomes and limited therapeutic options3. Understanding the molecular mechanisms driving disease progression and resistance is therefore crucial for identifying novel therapeutic targets and improving patient outcomes.
The development of preclinical models that accurately mimic the complexity of human prostate cancer is critical for advancing research in this field. Traditional 2D cell cultures, though widely used, fail to recapitulate the cellular heterogeneity, tumor architecture, and microenvironment observed in vivo4. As a result, the translational utility of these models is limited, particularly for studying tumor progression, drug resistance, and the dynamic interactions between cancer cells and the tumor microenvironment5. In contrast, patient-derived xenografts (PDXs) and organoid cultures have emerged as more sophisticated tools that bridge the gap between conventional cell lines and clinical tumors4,5,6.
Organoid culture systems have revolutionized cancer modeling. These three-dimensional (3D) structures are derived from patient tissues and can retain the histological, genomic, and phenotypic characteristics of the original tumor7,8,9,10.&#160;Importantly, prostate cancer organoids preserve critical features of disease heterogeneity, including AR signaling, resistance to treatment, and genomic instability, making them ideal tools for translational research11. Compared to PDX models, organoids are more cost-effective, scalable, and amenable to genetic manipulation, which facilitates high-throughput drug screening, biomarker discovery, and functional studies. Organoid platforms have already shown promising results in other malignancies, including colorectal, breast, and pancreatic cancers, underscoring their potential for personalized oncology4,12.
However, establishing and maintaining prostate cancer organoids (PCOs) presents unique technical challenges. The prostate epithelium is inherently dependent on androgen signaling and extracellular matrix (ECM) support, necessitating specialized culture conditions. Moreover, prostate cancer exhibits a wide range of clinical behaviors, from localized hormone-sensitive tumors to highly aggressive, treatment-refractory CRPC, which further complicates the development of reproducible organoid systems11,12. Successful protocols for PCO generation must address these biological complexities by optimizing key steps, including tissue processing, enzymatic digestion, ECM embedding, and supplementation with critical growth factors, hormones, and small molecules. Such protocols are essential for ensuring the fidelity of organoids to their parental tumors and facilitating their application in downstream analyses.
In this study, we present a detailed and reproducible protocol for the establishment and long-term maintenance of PDOs. Our workflow spans the entire process, from tissue acquisition and processing of biopsy and surgical specimens to organoid passaging, cryopreservation, and resuscitation (Figure 1). This protocol has been optimized to preserve the phenotypic and genomic integrity of prostate cancer tissues, enabling robust 3D culture systems suitable for a wide range of applications, including genomic profiling, drug response testing, and functional assays. Notably, our method addresses common technical hurdles, such as low success rates in organoid culture, suboptimal growth, and difficulties in subsequent passaging, and provides standardized guidelines, thereby facilitating reproducibility across laboratories.
The development of reliable prostate cancer organoid models holds immense significance for both basic and translational research13. These systems offer a powerful platform for deciphering disease mechanisms, identifying novel therapeutic targets, and advancing precision medicine approaches tailored to individual patients. By addressing existing technical gaps and providing a comprehensive protocol, our work aims to accelerate the adoption of organoid technology in prostate cancer research and support the global efforts to combat this challenging disease14.

<table><tbody><tr><td>A 83-01</td><td>TOCRIS</td><td>2939</td><td></td></tr><tr><td>adMEM/F12</td><td>Gibco</td><td>12634010</td><td></td></tr><tr><td>biosafety cabinet</td><td>Jinan Xinbeixi Biotechnology Co., Ltd</td><td></td><td></td></tr><tr><td>Cell Recovery Solution</td><td>Corning</td><td>354253</td><td></td></tr><tr><td>centrifuge</td><td>Shenzhen Ruiwode Life Technology Co., Ltd</td><td>M1416R</td><td></td></tr><tr><td>centrifuge tubes</td><td>Nanjing iResearch Biotechnology Co., Ltd</td><td></td><td></td></tr><tr><td>Cultrex PathClear BM</td><td>R&amp;amp;D</td><td>3533-010-02</td><td></td></tr><tr><td>Cultrex Reduced Growth Factor BME, Type 2</td><td>R&amp;amp;D</td><td>3533-005-02</td><td></td></tr><tr><td>culture containers (24-well low-adhesion plates)</td><td>thermo</td><td>Nunc-174930-ZX</td><td></td></tr><tr><td>DHT</td><td>ISOREAG</td><td>521-18-6</td><td></td></tr><tr><td>electronic balance</td><td>Nanjing iResearch Biotechnology Co., Ltd</td><td></td><td></td></tr><tr><td>Fetal Bovine Serum</td><td>Gibco</td><td>10099141</td><td></td></tr><tr><td>GlutaminePlus-200mM</td><td>R&amp;amp;D</td><td>B90210</td><td></td></tr><tr><td>HEPES</td><td>R&amp;amp;D</td><td>1254/10</td><td></td></tr><tr><td>humidified cell culture incubator (37&amp;deg;C, 5% CO2)</td><td>Shanghai Hetian Scientific Instrument Co., Ltd</td><td>HY-160SY</td><td></td></tr><tr><td>ice box</td><td>Nanjing iResearch Biotechnology Co., Ltd</td><td></td><td></td></tr><tr><td>medium-sized culture dishes</td><td>absin</td><td>abs7005</td><td></td></tr><tr><td>N21-MAX Media Supplement (50X)</td><td>R&amp;amp;D</td><td>AR008</td><td></td></tr><tr><td>N-Acetylcysteine</td><td>Sigma</td><td>A9165-5G</td><td></td></tr><tr><td>Nicotinamide</td><td>R&amp;amp;D</td><td>3533-010-02</td><td></td></tr><tr><td>Normocin</td><td>Invivogen</td><td>ant-nr-2</td><td></td></tr><tr><td>Pipet-aid pipetting device</td><td>Nanjing Hotspot Scientific Instrument Co., Ltd</td><td></td><td></td></tr><tr><td>pipettes</td><td>Nanjing Hotspot Scientific Instrument Co., Ltd</td><td></td><td></td></tr><tr><td>Primocin</td><td>Invivogen</td><td>ant-pm-2</td><td></td></tr><tr><td>rhEGF</td><td>R&amp;amp;D</td><td>236-EG-01M</td><td></td></tr><tr><td>rhFGF</td><td>R&amp;amp;D</td><td>233-FB-025</td><td></td></tr><tr><td>rhFGF-10</td><td>R&amp;amp;D</td><td>345-FG-025</td><td></td></tr><tr><td>rhNoggin</td><td>R&amp;amp;D</td><td>6057-NG-100</td><td></td></tr><tr><td>rhR-Spondin 1</td><td>R&amp;amp;D</td><td>4656-RS-100</td><td></td></tr><tr><td>SB 202190</td><td>TOCRIS</td><td>1264</td><td></td></tr><tr><td>Single-cell suspension preparation apparatus</td><td>RWD</td><td>DSC-400</td><td></td></tr><tr><td>Tissue scissors/scalpel</td><td>Nanjing iResearch Biotechnology Co., Ltd</td><td></td><td></td></tr><tr><td>TryplE Express</td><td>Gibco</td><td>126A04013</td><td></td></tr><tr><td>Tumor Dissociation Kit, Human</td><td>Miltenyi Biotec</td><td>130-095-929</td><td></td></tr><tr><td>tweezers</td><td>Nanjing iResearch Biotechnology Co., Ltd</td><td></td><td></td></tr><tr><td>water bath</td><td>Changzhou Zhongjie Experimental Instrument Manufacturing Co., Ltd</td><td>HH-600</td><td></td></tr><tr><td>Y-27632 dihydrochloride</td><td>R&amp;amp;D</td><td>1254/10</td><td></td></tr></tbody></table>

establishment and maintenance of patient-derived prostate cancer organoids: a detailed experimental protocol

Establishment and Maintenance of Patient-derived Prostate Cancer Organoids: A Detailed Experimental Protocol

Prostate cancer (PCa) is a highly heterogeneous malignancy and a leading cause of cancer-related mortality in men, with significant clinical challenges in addressing treatment resistance and disease progression. Existing preclinical models, such as two-dimensional (2D) cell lines and patient-derived xenografts (PDXs), have limitations in faithfully recapitulating the complexity of PCa, including tumor heterogeneity, androgen receptor (AR)-dependent signaling, and microenvironmental interactions. To address this gap, this study presents a robust and reproducible protocol for establishing Patient-derived organoids (PDOs) that preserve the genetic, phenotypic, and histological features of the parental tumors. This method enables the culture of organoids from both localized and advanced PCa, providing a biologically relevant model for studying disease mechanisms, drug response, and biomarker discovery. The results demonstrate the protocol's ability to generate organoids from a variety of clinical specimens, with applications in high-throughput drug screening and precision medicine. By offering a standardized workflow that addresses key technical challenges, this study highlights the importance of PDOs as versatile tools for advancing prostate cancer research and developing more effective therapeutic strategies.

Morphological and dynamic growth features of patient-derived prostate cancer organoids 
Patient-derived prostate cancer organoids were successfully established from fresh tumor tissues collected during clinical procedures. Initial organoid formation was typically observed within 5-7 days of culture (Figure 2). Under brightfield microscopy, organoids exhibited distinct morphological features, forming compact, well-defined spheroids or glandular-like structures. Organoids d...

Prostate cancer heterogeneity and treatment resistance remain challenges. We describe a protocol for generating patient-derived organoids (PDOs) that preserve key parental tumor characteristics. This standardized model supports drug screening and precision medicine advancements.

Prostate cancer (PCa) is a highly heterogeneous malignancy and a leading cause of cancer-related mortality in men, with significant clinical challenges in ...

The research focus on establishing and maintain the patient-derived prostate cancer organoid for personalized treatment studies. Advances include the optimized organoid culture system to augment the fidelity preservation and drug screening for personalized prostate cancer therapy. To begin place, collected tissue blocks into 50 milliliter sterile conical centrifuge tubes and label each tube accordingly.Add preservation solution to each tube until the tissues are fully submerged. Using sterilized tweezers, transfer the prostate cancer tissue samples into a medium-sized culture dish. Wash the tissues two to three times with two milliliters of DPBS to remove blood and debris.Then, carefully remove adipose tissue, muscle, and other non-epithelial components using fine-tipped tweezers, keeping only epithelial regions. Using scissors, mince the cleaned tissue into small fragments measuring approximately one to three cubic millimeters. Now, transfer the tissue fragments into tissue processing tubes and add tumor tissue digestion solution to each tube.Start the digestion device and run the medium-hard tissue dissociation program for enzymatic mechanical dissociation. After that, add an equal or double volume of FBS-supplemented adDMEM and F12 medium to the cell suspension to terminate enzymatic digestion. Pre-wet a 100 micrometer cell strainer with one milliliter of adDMEM/F12 medium and filter the cell suspension through it.Then, rinse the tissue processing tube and cell strainer twice with five milliliters of adDMEM/F12 medium supplemented with ROCK inhibitor solution. Collect the filtrate into a 50 milliliter sterile conical centrifuge tube. Pass the collected suspension through a 70 micrometer strainer and transfer all filtered suspension into a new 50 milliliter tube.Then, centrifuge the collected cell suspension at 200 G for five minutes at 25 degrees Celsius. Carefully discard the supernatant and resuspend the cell pellet in two milliliters of red blood cell lysis buffer. Then, incubate the suspension on ice for two to three minutes.To terminate the lysis, add six milliliters of adDMEM/F12 medium supplemented with ROCK inhibitor. Transfer the mixture to a 15 milliliter sterile conical centrifuge tube and centrifuge at 200 G for five minutes at 25 degrees Celsius. After discarding the supernatant, wash the cell pellet one to two times with the same supplemented medium and transfer the final suspension into a 1.5 milliliter Eppendorf tube.Now, mix 10 microliters of the cell suspension with 10 microliters of trypan blue solution. Resuspend the cell pellet in one milliliter of adDMEM/F12 medium. Pipette six microliters of the diluted suspension onto a hemo cytometer and count the cells under a microscope.Now, centrifuge the cell suspension at 200 G for five minutes at 25 degrees Celsius. Transfer the final cell pellet into a 1.5 milliliter Eppendorf tube. Determine the required volume of matrix gel from the cell count.Using pre-chilled pipette tips, carefully add the matrix gel to the cell pellet. Mix the cell gel mixture gently on ice to avoid air bubbles and maintain matrix gel integrity. Now, dispense approximately 30 microliters of the cell matrix gel mixture per drop into a low adhesion, 24-well culture plate.Place the culture plate in a humidified incubator at 37 degrees Celsius with 5%carbon dioxide for five minutes to allow preliminary gel solidification. Invert the plate and let it sit for 20 minutes to retain the three-dimensional form of the gel. Add 800 to 1, 000 microliters of culture medium to each well.Using a pre-chilled pipette tip, mechanically disrupt the matrix gel in each well to fragment the organoid clusters. Rinse each well one to two times with adDMEM/F12 medium supplemented with ROCK inhibitor solution. Transfer the fragmented clusters and culture medium into a 15 milliliter sterile conical centrifuge tube.Centrifuge the tube at 200 G for five minutes at 25 degrees Celsius. After discarding the supernatant, resuspend the resulting pellet in two milliliters of pre-warmed recombinant trypsin-like enzymes solution. Pipette the mixture up and down to begin dissociation.Then, incubate the tube at 37 degrees Celsius for 15 to 30 minutes. Now, add an equal or double volume of adDMEM/F12 medium supplemented with FBS to stop the enzymatic reaction. Centrifuge the suspension at 200 G for five minutes at 25 degrees Celsius.Then, discard the supernatant. To begin matrix gel removal, resuspend the pellet in two milliliters of cell recovery solution. Incubate the suspension at four degrees Celsius for 20 to 40 minutes to dissolve the residual matrix gel.Centrifuge the sample at 200 G for five minutes at four degrees Celsius. After discarding the supernatant, wash the pellet two times using adDMEM and F12 medium to eliminate any remaining matrix components. Finally, transfer the pellet to a 1.5 milliliter sterile Eppendorf tube for further processing.Seed the dissociated organoid clusters following the reseeding procedures. Initial organoid formation was observed between day five and day seven of culture with bright-field images showing compact and well-defined spheroids or glandular-like structures. Hematoxylin and eosin staining revealed preserved glandular architecture in organoids derived from well-differentiated adenocarcinoma, resembling the matched patient tissue.Immunohistochemistry showed strong nuclear androgen receptor expression in both organoids and primary tumors derived from hormone-sensitive prostate cancer tissues. CK5, PSMA, and NKX3.1 were robustly expressed in both organoid and tumor samples, supporting epithelial origin and lineage fidelity. KI-67 staining showed that the organoids retained a proliferative index comparable to the original tumor tissues.We have developed a robust protocols for maintaining patient-derived prostate cancer organoids, enabling personalized drug testing and biomarker discovery. Our findings provide a reliable platform for studying prostate cancer biology, facilitating personalized therapies and precision medicine advancements. Future research will explore organoid heterogeneity, tumor microenvironment interactions, and drug resistance mechanisms in prostate cancer.

establishment-maintenance-patient-derived-prostate-cancer-organoids

Research

JoVE Journal

Cancer Research

Organoids derived from tumors are a type of 3D cell culture model that can maintain the pathological features and organ-specific cell lineage of tumors. In the example protocol, we will see scientists generate organoids from a prostate tumor from a genetically engineered mouse model.

Preparing Organoids from Tumors: A 3D In Vitro Tumor Model

Here, we present a protocol to guide human primary prostate organoid handling then suggest endpoints to assess phenotype. Seeding, culture maintenance, recovery from matrix gel, morphologic quantification, embedding and sectioning, FFPE sectioning, whole-mount staining, and application of commercial assays are described.

Handling and Assessment of Human Primary Prostate Organoid Culture

We show a method for necropsy and dissection of mouse prostate cancer models, focusing on prostate tumor dissection. A step-by-step protocol for generation of mouse prostate tumor organoids is also presented.

Generation of Tumor Organoids from Genetically Engineered Mouse Models of Prostate Cancer

Presented here is a protocol to study pharmacological responses in prostate epithelial organoids. Organoids closely resemble in vivo biology and recapitulate patient genetics, making them attractive model systems. Prostate organoids can be established from wildtype prostates, genetically engineered mouse models, benign human tissue, and advanced prostate cancer.

Prostate Organoid Cultures as Tools to Translate Genotypes and Mutational Profiles to Pharmacological Responses

Three-dimensional cultures of patient BMPC specimens and xenografts of bone metastatic prostate cancer maintain the functional heterogeneity of their original tumors resulting in cysts, spheroids and complex, tumor-like organoids. This manuscript provides an optimization strategy and protocol for 3D culture of heterogeneous patient derived samples and their analysis using IFC.

Establishment and Analysis of Three-Dimensional (3D) Organoids Derived from Patient Prostate Cancer Bone Metastasis Specimens and their Xenografts

A detailed protocol is provided here for establishing human breast organoids from patient-derived breast tumor resections or normal breast tissue. The protocol provides comprehensive step-by-step instructions for culturing, freezing, and thawing human patient-derived breast organoids.

Establishment and Culture of Patient-Derived Breast Organoids

Tumor organoids have revolutionized cancer research and the approach to personalized medicine. They represent a clinically relevant tumor model that allows researchers to stay one step ahead of the tumor in the clinic. This protocol establishes tumor organoids from fresh pancreatic tumor tissue samples and patient-derived xenografts of pancreatic adenocarcinoma origin.

Establishment of Pancreatic Cancer-Derived Tumor Organoids and Fibroblasts From Fresh Tissue

This protocol describes the generation of a murine prostate organoid-based model grown in 2D. It includes organoid dissociation, cell seeding, differentiation, epithelial integrity assessment, and in-vitro infection with uropathogenic Escherichia coli. The model provides apical accessibility and is suitable for studying host-pathogen interactions, overcoming limitations of traditional 3D organoid systems.

Generation of a Mouse Prostate Organoid-Based Model for Studying Host-Pathogen Interactions

Utilizing patient-derived tumors in a subcutaneous preclinical model is an excellent way to study the efficacy of novel therapies, predictive biomarker discovery, and drug resistant pathways. This model, in the drug development process, is essential in determining the fate of many novel anti-cancer therapies prior to clinical investigation.

Development and Maintenance of a Preclinical Patient Derived Tumor Xenograft Model for the Investigation of Novel Anti-Cancer Therapies

The following protocol focuses on the establishment of a primary culture of patient-derived soft tissue sarcoma (STS). This model could help us to better understand the molecular background and pharmacological profile of these rare malignancies and could represent a starting point for further research aimed at improving STS management.

Establishment of a Primary Culture of Patient-derived Soft Tissue Sarcoma

Learn to establish patient-derived prostate cancer organoids using optimized culture protocols. This method preserves tumor fidelity for personalized drug screening and precision medicine research.

Establishment and Maintenance of Patient-derived Prostate Cancer Organoids: A Detailed Experimental Protocol

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