【AI Explained】Organoid culture systems for prostate epithelial and cancer tissue.

Literature Information

TL;DR

This study presents a protocol for creating 3D prostate organoid cultures from healthy and metastatic prostate cancer tissues, which successfully replicate the cellular architecture of both normal and cancerous prostate cells. The ability to grow organoids from various prostate cell types in a defined serum-free medium not only advances the understanding of prostate biology and tumorigenesis but also facilitates drug discovery efforts targeting prostate cancer.

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Organoid culture · Prostate cancer · Tumor biology

Abstract

This protocol describes a strategy for the generation of 3D prostate organoid cultures from healthy mouse and human prostate cells (either bulk or FACS-sorted single luminal and basal cells), metastatic prostate cancer lesions and circulating tumor cells. Organoids derived from healthy material contain the differentiated luminal and basal cell types, whereas organoids derived from prostate cancer tissue mimic the histology of the tumor. We explain how to establish these cultures in the fully defined serum-free conditioned medium that is required to sustain organoid growth. Starting with the plating of digested tissue material, full-grown organoids can usually be obtained in ∼2 weeks. The culture protocol we describe here is currently the only one that allows the growth of both the luminal and basal prostatic epithelial lineages, as well as the growth of advanced prostate cancers. Organoids established using this protocol can be used to study many different aspects of prostate biology, including homeostasis, tumorigenesis and drug discovery.

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Primary Questions Addressed

1. What are the specific advantages of using 3D organoid cultures over traditional 2D cell cultures in studying prostate cancer?
2. How do the characteristics of organoids derived from healthy prostate tissue differ from those derived from metastatic prostate cancer lesions?
3. What role do circulating tumor cells play in the development of organoid cultures for prostate cancer research?
4. Can the protocol described for organoid culture be adapted for other types of cancer, and if so, how?
5. What are the potential applications of prostate organoids in personalized medicine and drug discovery for prostate cancer patients?

Key Findings

Research Summary: Organoid Culture Systems for Prostate Epithelial Tissue and Prostate Cancer Tissue

Background and Purpose

The study introduces a protocol for generating three-dimensional (3D) organoid cultures from both healthy and cancerous prostate tissues. This method aims to overcome limitations in existing in vitro models, particularly in studying prostate biology, tumorigenesis, and drug discovery. The protocol is designed to create organoids that accurately reflect the cellular architecture and signaling pathways of prostate tissue.

Main Methods/Materials/Experimental Design

The authors developed a detailed stepwise protocol for establishing organoid cultures from mouse and human prostate tissues, including metastatic lesions and circulating tumor cells. Key components of the protocol include:

1. Study Design: The study employs a systematic approach to establish and maintain organoid cultures, focusing on both healthy and cancerous prostate tissues.

2. Recruitment Criteria:

   • Healthy mouse prostate tissue and human prostate samples (both healthy and cancerous).
   • Inclusion of metastatic biopsy samples and circulating tumor cells from patients.

3. Organoid Culture Methodology:

   • Tissues are minced and digested using collagenase and TrypLE.
   • The resultant cell suspension is mixed with Matrigel and cultured in defined serum-free medium containing growth factors.
   • Specific conditions are optimized for both mouse and human prostate organoids.

4. Endpoints:

   • Primary endpoints include the successful establishment and growth of organoids.
   • Secondary endpoints involve the characterization of organoids to ensure they reflect the original tissue’s phenotype and genotype.

5. Statistical Analysis: The efficiency of organoid formation and growth is assessed qualitatively and quantitatively, ensuring reproducibility and reliability of the results.

Key Results and Findings

• The protocol achieved a high efficiency of over 95% for establishing organoid cultures from healthy prostate tissues, while the efficiency dropped to 15-20% for advanced prostate cancer tissues.
• Organoids derived from healthy tissues displayed both luminal and basal cell types, mimicking the histological features of prostate tissue.
• Prostate cancer organoids maintained genetic and phenotypic characteristics reflective of their original tumors, providing a valuable model for cancer research.

Conclusion/Significance/Innovation

This research presents a significant advancement in prostate cancer modeling by providing a reliable method for generating organoids that can be used for various applications, including drug discovery and understanding prostate biology. The use of organoids offers a more physiologically relevant model compared to traditional cell lines, potentially leading to better therapeutic strategies.

Research Limitations and Future Directions

• The study acknowledges that while the organoid culture system is promising, challenges remain in obtaining sufficient material from advanced cancer tissues.
• Future research could focus on improving the efficiency of organoid establishment from cancerous tissues and exploring the genetic modifications of organoids to better study specific oncogenic pathways.
• The authors suggest further investigations into the heterogeneity of organoid cultures derived from different patients to enhance personalized medicine approaches in prostate cancer treatment.

References

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Literatures Citing This Work

1. Prostate epithelial cell of origin determines cancer differentiation state in an organoid transformation assay. - Jung Wook Park;John K Lee;John W Phillips;Patrick Huang;Donghui Cheng;Jiaoti Huang;Owen N Witte - Proceedings of the National Academy of Sciences of the United States of America (2016)
2. Assay Establishment and Validation of a High-Throughput Screening Platform for Three-Dimensional Patient-Derived Colon Cancer Organoid Cultures. - Karsten Boehnke;Philip W Iversen;Dirk Schumacher;María José Lallena;Rubén Haro;Joaquín Amat;Johannes Haybaeck;Sandra Liebs;Martin Lange;Reinhold Schäfer;Christian R A Regenbrecht;Christoph Reinhard;Juan A Velasco - Journal of biomolecular screening (2016)
3. Major milestones in translational oncology. - Tommaso A Dragani;Antoni Castells;Vathany Kulasingam;Eleftherios P Diamandis;Helena Earl;Wade T Iams;Christine M Lovly;J P Michiel Sedelaar;Jack A Schalken - BMC medicine (2016)
4. Wnt Drug Discovery: Weaving Through the Screens, Patents and Clinical Trials. - Benjamin Lu;Brooke A Green;Jacqueline M Farr;Flávia C M Lopes;Terence J Van Raay - Cancers (2016)
5. Impact of label-free technologies in head and neck cancer circulating tumour cells. - Arutha Kulasinghe;Liz Kenny;Chris Perry;Jean-Paul Thiery;Lidija Jovanovic;Ian Vela;Colleen Nelson;Chamindie Punyadeera - Oncotarget (2016)
6. Bioluminescence Microscopy as a Method to Measure Single Cell Androgen Receptor Activity Heterogeneous Responses to Antiandrogens. - Pallavi Jain;Bertrand Neveu;Lauriane Velot;Lily Wu;Yves Fradet;Frédéric Pouliot - Scientific reports (2016)
7. Efficient and simple approach to in vitro culture of primary epithelial cancer cells. - Karolina Janik;Marta Popeda;Joanna Peciak;Kamila Rosiak;Maciej Smolarz;Cezary Treda;Piotr Rieske;Ewelina Stoczynska-Fidelus;Magdalena Ksiazkiewicz - Bioscience reports (2016)
8. Conditionally reprogrammed normal and primary tumor prostate epithelial cells: a novel patient-derived cell model for studies of human prostate cancer. - Olga A Timofeeva;Nancy Palechor-Ceron;Guanglei Li;Hang Yuan;Ewa Krawczyk;Xiaogang Zhong;Geng Liu;Geeta Upadhyay;Aleksandra Dakic;Songtao Yu;Shuang Fang;Sujata Choudhury;Xueping Zhang;Andrew Ju;Myeong-Seon Lee;Han C Dan;Youngmi Ji;Yong Hou;Yun-Ling Zheng;Chris Albanese;Johng Rhim;Richard Schlegel;Anatoly Dritschilo;Xuefeng Liu - Oncotarget (2017)
9. Dissecting cell-type-specific roles of androgen receptor in prostate homeostasis and regeneration through lineage tracing. - Qing Xie;Yueli Liu;Tao Cai;Corrigan Horton;Joshua Stefanson;Zhu A Wang - Nature communications (2017)
10. A Tale of Two Signals: AR and WNT in Development and Tumorigenesis of Prostate and Mammary Gland. - Hubert Pakula;Dongxi Xiang;Zhe Li - Cancers (2017)

… (376 more literatures)

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