Endometrial Organoids in PCOS: A Translational Tool for Gynecologic Precision Medicine

Arwa Ibrahim

doi:10.20944/preprints202506.2488.v1

Submitted:

27 June 2025

Posted:

30 June 2025

You are already at the latest version

Abstract

Background:Polycystic ovary syndrome (PCOS) is among the most common endocrine disorders affecting women of reproductive age and is closely linked to endometrial dysfunction and fertility challenges. Gaining a clearer understanding of these changes is essential for improving clinical care and outcomes.Objective:This mini-review highlights recent developments in using endometrium-derived epithelial organoids to model PCOS-related conditions, with a focus on their translational and clinical significance.Methods:A focused review of recent literature was conducted on PCOS-specific endometrial organoids and their application in disease modeling, drug testing, and tissue engineering. Emphasis was placed on studies highlighting molecular characterization and regenerative strategies.Results:Organoids derived from PCOS endometrium have shed light on hormonal imbalances, gene expression shifts, and cellular dysfunctions linked to the condition. These 3D models effectively mimic endometrial pathology and are being used to explore potential treatments. Additionally, efforts to integrate organoids with biomaterials are paving the way for regenerative solutions.Conclusion:Endometrial organoids represent a powerful and clinically meaningful platform for studying PCOS-related endometrial changes. Their growing role in personalized and regenerative medicine holds significant promise for tackling infertility and other reproductive health issues in PCOS.

Keywords:

PCOS

;

endometrial organoids

;

tissue engineering

;

epithelial organoids

;

endometrial dysfunction

Subject:

Medicine and Pharmacology - Other

Introduction

One of the most common endocrine conditions affecting women of reproductive age is polycystic ovary syndrome (PCOS), which is characterized by polycystic ovarian morphology, hyperandrogenism, and ovulatory failure [1]. Because of progesterone resistance and prolonged estrogen exposure, the endometrium in PCOS women has unique pathological characteristics, such as impaired decidualization, altered receptivity patterns, and increased proliferative activity. Traditional two-dimensional cell culture models have proven inadequate for studying the complex cellular interactions and hormonal responses characteristic of PCOS endometrium [2,3,4].

A paradigm change in reproductive biology research has been made possible by the emergence of three-dimensional organoid technology, which has made it possible to create physiologically accurate models that recapitulate tissue architecture and function [5,6]. Endometrial epithelial organoids created from PCOS patients provide special chances to investigate disease-specific processes while preserving the pathogenic characteristics of the original tissue [7]. These models show great promise for diagnosing endometrial malfunction, identifying therapeutic approaches, and creating individualized treatment plans.

PCOS Endometrial Pathophysiology and Organoid Development

Cellular and Molecular Alterations in PCOS Endometrium

Comprehensive cellular atlases of the PCOS endometrium have been found by single-cell RNA sequencing studies [3,8], which show notable changes in the composition of cells and patterns of gene expression. Compared to healthy controls, PCOS women's proliferative phase endometrium has lower populations of stromal cells and higher proportions of epithelial cells. Cell-type-specific disease signatures that impact metabolic processes, hormone responsiveness, and inflammatory pathways accompany these compositional changes [8,9].

Through androgen receptor-mediated signaling, hyperandrogenism, a defining feature of PCOS, directly affects the cellular activity of the endometrium [10,11]. Overproduction of androgens can raise the risk of endometrial cancer in PCOS patients by promoting cellular proliferation and dysregulating gene expression patterns. Organoid systems obtained from patients can be used to adequately replicate the distinct endometrial microenvironment created by prolonged exposure to increased androgen levels [12].

Establishment of PCOS Endometrial Organoids

The successful establishment of PCOS-derived endometrial epithelial organoids marks a significant technological breakthrough in the study of reproductive biology [7]. Enzymatic dissociation techniques are used to generate these organoids from endometrial biopsies taken during the proliferative phase of the menstrual cycle, which are then cultured in three dimensions in extracellular matrix scaffolds [6,7,13]. The organoids exhibit hormone response comparable to original tissue and preserve the integrity of the proliferative basolateral epithelial membrane [15].

When compared to control organoids, PCOS organoids show unique traits such as differential gene expression patterns linked to inflammation and receptivity and changed responses to sex hormone exposure [7]. Importantly, these models maintain disease-specific traits even after several passes, which makes them useful tools for therapeutic screening applications and longitudinal studies.

Tissue Engineering Applications and Therapeutic Potential

Bioengineering Approaches for Endometrial Repair

New treatment options for endometrial dysfunction have been made possible by the combination of tissue engineering techniques with organoid technology. In order to create complex three-dimensional structures that more closely resemble natural tissue architecture, vascularized endometrial organoids were created by co-culturing epithelial organoids with endometrial stromal cells and human umbilical vein endothelial cells [16,17]. These engineered structures show improved therapeutic potential for endometrial regeneration and repair.

By making it possible to precisely control hormone gradients and cellular microenvironments, organ-on-chip technologies and microfluidic devices have significantly advanced the field [18,19,20]. These platforms enable drug response studies under physiologically appropriate settings and dynamic simulation of menstrual cycle stages. The pathophysiology of PCOS has been better understood because to research on the bidirectional endocrine crosstalk between the ovary and endometrium made possible by the development of dual reproductive organ-on-chip systems [18,19,20].

Drug Screening and Personalized Medicine

Patient-derived PCOS organoids serve effective tools for drugs screening and the creation of tailored treatment strategies. These models have been used to assess how metformin therapy affects the endometrium of PCOS, showing decreased expression of androgen receptors and restoration of normal gene expression patterns [11]. Rapid evaluation of medicinal substances and the discovery of new therapy targets are made possible by high-throughput screening techniques employing organoid models. Studying treatment responses in a variety of patient groups is made possible by the capacity to create organoid biobanks from PCOS patients [3,7,11]. After thawing, cryopreserved organoids retain their characteristics [21]. This method makes it easier to create precision medicine plans that are specific to each patient's characteristics and disease manifestations. The following table provides a comparative overview of various PCOS endometrial organoid models, outlining their culture systems, distinguishing features, and specific applications in drug screening and personalized medicine, along with their respective advantages and limitations.

Table 1. Comparison of PCOS Endometrial Organoid Models and Applications [3,7,12,17,18,19,20].

Model Type	Culture System	Key Features	Applications	Advantages	Limitations
Scaffold-free organoids	Matrigel-embedded	Epithelial-stromal organization, hormone responsiveness	Androgen response studies, cancer risk assessment	Physiological architecture, long-term culture	Limited vascularization
Epithelial organoids	3D Matrigel culture	Pure epithelial population, disease-specific signatures	Hormone response screening, biomarker discovery	Disease trait retention, high reproducibility	Lacks stromal interactions
Vascularized organoids	Co-culture system	Epithelial-stromal-endothelial integration	Therapeutic screening, regenerative applications	Enhanced physiological relevance	Technical complexity
Microfluidic models	Organ-on-chip platform	Dynamic hormone exposure, controlled gradients	Cycle modeling, drug testing	Precise control, real-time monitoring	Limited throughput
Assembloid systems	Multi-cellular co-culture	Epithelial-stromal interactions, implantation modeling	Fertility research, therapeutic development	Comprehensive cellular interactions	Variable reproducibility

Challenges and Future Directions

Technical and Methodological Considerations

Despite significant advances, several challenges remain in PCOS organoid research and clinical translation . Standardization of culture protocols, matrix compositions, and assessment methods is critical for ensuring reproducibility across laboratories [7,22]. The replacement of animal-derived matrices with synthetic alternatives represents an important step toward clinical applications. Therapeutic screening applications have trouble with organoid heterogeneity and batch-to-batch variability [23]. For therapeutic screening applications, organoid heterogeneity and batch-to-batch variability present challenges. To create robust quality standards, advanced quality control techniques are required, such as multi-omics characterisation and standardized functional assays. For clinical applications, automating organoid culture procedures may aid increase scalability and decrease variability [7,12,24].

Clinical Translation and Regulatory Considerations

Integration of organoid data with clinical parameters, biomarker profiles, and treatment responses will be essential for establishing the clinical relevance of these models [25]. Regulatory frameworks for organoid-based drug screening and personalized medicine approaches are still evolving and require continued development. Future directions involve establishing multi-center collaborative networks for organoid research, integrating artificial intelligence for pattern recognition and drug discovery, and creating more sophisticated co-culture systems with immune cells [25]. Studying causal links and creating focused therapeutic treatments are made possible by the combination of gene editing techniques with organoid technology.

Conclusions

An innovative technique for understanding endometrial dysfunction and creating individualized treatment plans is PCOS endometrium-derived epithelial organoids. These models offer platforms for examining hormone responses, inflammatory processes, and therapeutic treatments in addition to successfully capturing disease-specific molecular fingerprints. Regenerative medicine and sophisticated drug screening platforms are among the many potential uses for organoid technology that have been made possible by its integration with tissue engineering techniques. The ongoing development of PCOS organoid models has great potential for advancing our knowledge of endometrial pathophysiology and creating more effective treatments for affected women, even though standardization and clinical translation still present difficulties. The development of cooperative research networks and organoid biobanks will accelerate the shift to personalized medicine strategies for PCOS treatment. To take full advantage of the therapeutic potential of this technology, future research should concentrate on increasing model complexity, refining standardization processes, and confirming clinical applications.

Acknowledgments

During the preparation of this work the author used [Perplexity] in order to [Rephrase already written sentences]. After using this tool, the author reviewed and edited the content as needed and takes full responsibility for the content of the publication.

Conflicts of Interest

The author declares no conflict of interest.

References

Jiawen, D., & Rees, D. A. (2023). Polycystic ovary syndrome: Pathophysiology and therapeutic opportunities. BMJ Medicine, 2(1). [CrossRef]
Vanden Brink, H., Pisch, A. J., & Lujan, M. E. (2021). A comparison of two-and three-dimensional ultrasonographic methods for evaluation of ovarian follicle counts and classification of polycystic ovarian morphology. Fertility and Sterility, 115(3), 761-770.
Eriksson, G., Mansouri, M. R., Martínez-Zamora, M. A., Papadogiannakis, N., Elenis, E., Gidlöf, S., Stavreus-Evers, A., Hirschberg, A. L., & Stener-Victorin, E. (2025). Single-cell profiling of the human endometrium in polycystic ovary syndrome. Nature Medicine, 31(7), 1925-1938. [CrossRef]
Kim, J., Koo, B. K., & Knoblich, J. A. (2020). Human organoids: Model systems for human biology and medicine. Nature Reviews Molecular Cell Biology, 21(10), 571-584. [CrossRef]
Guo J, Zhou W, Sacco M, Downing P, Dimitriadis E, Zhao F. Using organoids to investigate human endometrial receptivity. Front Endocrinol (Lausanne). 2023 Aug 24;14:1158515. [CrossRef]
Boretto M, Cox B, Noben M, Hendriks N, Fassbender A, Roose H, Amant F, Timmerman D, Tomassetti C, Vanhie A, Meuleman C, Ferrante M, Vankelecom H. Development of organoids from mouse and human endometrium showing endometrial epithelium physiology and long-term expandability. Development. 2017 May 15;144(10):1775-1786. [CrossRef]
Luyckx L, Wei M, Saarela U, Myllykangas M, Kinnunen J, Arffman R, Lie Fong S, Vriens J, Vankelecom H, Piltonen TT. PCOS endometrium-derived epithelial organoids as a novel model to study endometrial dysfunction. Hum Reprod. 2025 Jun 11:deaf113. [CrossRef]
A cell atlas of human endometrium reveals key molecular and cellular alterations in PCOS. Nat Med 31, 1763–1764 (2025). [CrossRef]
Eriksson, G., Li, C., Han-Pin, P., Risal, S., Hirschberg, A. L., & Petropoulos, S. (2022). Endometrial cell-type-specific disease signatures and endometrial organoids in polycystic ovary syndrome. Endocrine Abstracts, 81, EP871. [CrossRef]
Abbey C Lissaman, Jane E Girling, Lynsey M Cree, Rebecca E Campbell, Anna P Ponnampalam, Androgen signalling in the ovaries and endometrium, Molecular Human Reproduction, Volume 29, Issue 6, June 2023, gaad017. [CrossRef]
Ohara, M., Yoshida-Komiya, H., Ono-Okutsu, M., Yamaguchi-Ito, A., Takahashi, T., & Fujimori, K. (2021). Metformin reduces androgen receptor and upregulates homeobox A10 expression in uterine endometrium in women with polycystic ovary syndrome. Reproductive Biology and Endocrinology, 19, 77. [CrossRef]
Wiwatpanit T, Murphy AR, Lu Z, Urbanek M, Burdette JE, Woodruff TK, Kim JJ. Scaffold-Free Endometrial Organoids Respond to Excess Androgens Associated With Polycystic Ovarian Syndrome. J Clin Endocrinol Metab. 2020 Mar 1;105(3):769–80. [CrossRef]
Dekanski, A., Eriksson, G., Turunen, T., Vesterinen, A., Li, C., Zheng, X., Gorsek-Sparovec, T., Piltonen, T., Hirschberg, A. L., Taebnia, N., & Stener-Victorin, E. (2025). Modelling polycystic ovary syndrome in vitro with endometrium epithelial organoids and stroma cell culture. Endocrine Abstracts, 110, P992. [CrossRef]
Eriksson G, Li C, Risal S, Pui HP, Torstensson S, Hirschberg AL, Petropoulos S, Deng Q, Stener-Victorin E. FRI393 Mapping Endometrial Cell-type-specific Disease Signatures A nd Endometrial Organoids In Polycystic Ovary Syndrome. J Endocr Soc. 2023 Oct 5;7(Suppl 1):bvad114.1587. [CrossRef]
Hernandez-Gordillo, V., Kassis, T., Lampejo, A., Choi, G., Gamboa, M. E., Gnecco, J. S., Brown, A., Breault, D. T., Carrier, R., & Griffith, L. G. (2020). Fully synthetic matrices for in vitro culture of primary human intestinal enteroids and endometrial organoids. Biomaterials, 254, 120125. [CrossRef]
Lloyd-Griffith, C., McFadden, T. M., Duffy, G. P., Unger, R. E., Kirkpatrick, C. J., & O’Brien, F. J. (2015). The pre-vascularisation of a collagen-chondroitin sulphate scaffold using human amniotic fluid-derived stem cells to enhance and stabilise endothelial cell-mediated vessel formation. Acta Biomaterialia, 26, 263–273. [CrossRef]
Y Dai, F Zhao, S Zhang, Q Chen, B Zeng, X Wang, W Gu, Y Zhang, X Lin, N Liu, F Zhou, J Yang, S Xin, Y Feng, P-790 Engineering Vascularized Human Endometrial Organoids for In Vivo Tissue Regeneration and Repair Applications, Human Reproduction, Volume 39, Issue Supplement_1, July 2024, deae108.054. [CrossRef]
An L, Liu Y, Liu Y. Organ-on-a-Chip Applications in Microfluidic Platforms. Micromachines (Basel). 2025 Feb 10;16(2):201. [CrossRef]
Campo H, Murphy A, Yildiz S, Woodruff T, Cervelló I, Kim JJ. Microphysiological Modeling of the Human Endometrium. Tissue Eng Part A. 2020 Jul;26(13-14):759-768. [CrossRef]
Park SR, Kim SR, Lee JW, Park CH, Yu WJ, Lee SJ, Chon SJ, Lee DH, Hong IS. Development of a novel dual reproductive organ on a chip: recapitulating bidirectional endocrine crosstalk between the uterine endometrium and the ovary. Biofabrication. 2020 Oct 16;13(1). [CrossRef]
Chen JC, Hoffman JR, Arora R, Perrone LA, Gonzalez-Gomez CJ, Vo KC, Laird DJ, Irwin JC, Giudice LC. Cryopreservation and recovery of human endometrial epithelial cells with high viability, purity, and functional fidelity. Fertil Steril. 2016 Feb;105(2):501-10.e1. [CrossRef]
Yaqing Wang, Jianhua Qin, Advances in human organoids-on-chips in biomedical research, Life Medicine, Volume 2, Issue 1, February 2023, lnad007. [CrossRef]
Gu Y, Zhang W, Wu X, Zhang Y, Xu K, Su J. Organoid assessment technologies. Clin Transl Med. 2023 Dec;13(12):e1499. [CrossRef]
EATRIS-Plus Multi-omics working group and Stakeholders. Multi-omics Quality Assessment in Personalized Medicine Through European Infrastructure for Translational Medicine (EATRIS): An Overview. Phenomics (2025). [CrossRef]
Haowei Yang, Jiawei Li, Zitian Wang, Davit Khutsishvili, Jiyuan Tang, Yu Zhu, Yongde Cai, Xiaoyong Dai, Shaohua Ma, Bridging the organoid translational gap: integrating standardization and micropatterning for drug screening in clinical and pharmaceutical medicine, Life Medicine, Volume 3, Issue 2, April 2024, lnae016. [CrossRef]

Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

© 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

Copyright: This open access article is published under a Creative Commons CC BY 4.0 license, which permit the free download, distribution, and reuse, provided that the author and preprint are cited in any reuse.