Emerging Frontiers in Ovarian Organoids: Bridging Development, Function, and Disease Modeling: A Review

Abstract

This review highlights recent advances in ovarian organoid research. Ovarian organoids are three-dimensional (3D) structures derived from primary ovarian tissues, cancer cells, or stem cells that replicate key architectural and func tional features of native ovarian tissue. Their formation requires both germ cells, such as oocytes and primordial germ cells, and somatic cells, including stromal, thecal, follicular, and epithelial cells. Ovarian organoids are typically characterized through histological, molecular, and functional analyses to confirm their structural and transcriptional resemblance to the native ovary. These organoids contain a heterogeneous population of cell types, reflecting the cel lular diversity of ovarian tissue, and exhibit gene expression profiles closely aligned with those of primary ovarian tissues. Organoids derived from both normal and malignant sources hold great potential for a wide range of applica tions, including basic ovarian biology, cancer research and therapeutics, fertility studies, drug screening, disease and cancer modeling, endocrine function studies, personalized medicine, and pathogen interaction analysis. Despite exist ing technical and biological challenges, ongoing research and innovations continue to expand the potential of ovarian organoids in reproductive biology and disease management.

Introduction

Advances in tissue engineering and the optimization of two-dimensional (2D) and three-dimensional (3D) culture systems have enabled researchers to structures that closely mimic the architecture and physiology of native tissues. These self-organizing structures, known as organoids, and were first reported in 1946 as cyst-like formations (1). Since then, organoid research has expanded across a wide range of tissues and organs, including the female reproductive tract, where studies demonstrated that adult stem cells can be used to recreate organ-like structures in vitro (2-4).

In this review, we summarize recent developments and emerging insights into ovarian organoids, focusing on their generation, characterization, and potential applications in reproductive biology and disease modeling.

Ovarian organoids

Ovarian organoids are 3D structures derived from primary ovarian tissues, cancer cells, or stem cells that recapitulate the key architectural and functional characteristics of native ovarian tissue, representing a major advancement in reproductive biology. This architecture results from the intrinsic ability of ovarian cells to self- organize when cultured within a supportive extracellular matrix (ECM) composed of natural or synthetic biomaterials.

Typically, ovarian organoids exhibit follicle-like structures, featuring a central oocyte surrounded by granulosa and theca cells, thereby mimicking the functional units of the ovary. In some models, stromal components—comprising connective tissue and supporting cells—are also represented, providing critical insights into intercellular interactions among diverse ovarian cell types, including mesenchymal stem cells and fibroblasts.

The complex 3D organization, cellular heterogeneity, functional responsiveness to hormonal and pharmacological stimuli, and physiologically relevant gene expression profiles of ovarian organoids make them powerful systems for studying ovarian biology, cancer progression, disease modeling, reproductive health, and personalized medicine. Compared with conventional 2D cultures, ovarian organoids offer a more realistic physiological context for investigating ovarian function and pathology (2, 3, 5). Further characterization of these models will deepen our understanding of ovarian physiology and contribute to the development of innovative therapeutic strategies for ovarian-related disorders.

Organoids are typically characterized using histological staining, gene expression profiling, and functional assays to validate their structural and transcriptional similarity to native ovarian tissues. They comprise a heterogeneous population of cell that mirrors the cellular diversity of the ovary. Moreover, ovarian organoids exhibit gene expression profiles closely aligned with primary ovarian tissues, including key markers such as follicle-stimulating hormone receptor (FSHR), luteinizing hormone/choriog onadotropin (LHCGR), anti-Müllerian hormone (AMH), growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), forkhead box L2 (FOXL2), and aromatase (CYP19A1).

A hallmark feature of ovarian organoids is their capacity to produce hormones similar to those of the native ovary. These organoids can synthesize and secrete estrogens and progesterone in response to hormonal stimuli, enabling the study of endocrine responses under both physiological and pathological conditions. This functionality is particularly valuable for investigating disorders such as polycystic ovary syndrome and hormone-dependent cancers. Overall, ovarian organoids provide a transformative platform for exploring ovarian biology and disease, offering a physiologically relevant alternative to conventional 2D cultures. Despite technical and biological challenges, ongoing research and technological innovations continue to expand their potential for advancing reproductive health and disease modeling.

Generation and sources of ovarian organoids

The ovary is a central organ of the female reproductive system, with follicles serving as its fundamental functional units. Each follicle comprises a germ cell, or oocyte, surrounded by somatic cells. The number and functional capacity of follicles change throughout life. Follicle formation begins before birth or shortly thereafter, followed by oocyte development and maturation, concurrent with the proliferation of follicular and theca cells and the secretion of hormones, including estrogen and progesterone. At birth, the ovary contains approximately 1-2 million germ cells, which gradually decline until puberty. Following regular menstrual cycles, this number continues to decrease, ultimately reaching around 1,000 at menopause.

Ovarian organoid generation requires both germ cells, such as oocytes and primordial germ cells, and somatic cells, including stromal, thecal, follicular, and ovarian epithelial cells. The various cell sources for ovarian organoid formation are summarized in Figure 1. These cells can be cultured using either scaffold-based or scaffold- free systems (6). Stromal cells can be isolated from fetal or adult ovarian tissue, and numerous studies have demonstrated the differentiation of pluripotent stem cells into stromal cells, as well as the derivation of germ cells or oocytes from pluripotent stem cells (7, 8). In this review, we focus on ovarian organoid formation from diverse cell types, including embryonic, fetal, and adult ovarian tissues, stem cells, and cancer-derived cell lines.

Fig.1.

Cell sources for ovarian organoid generation. MSCs; Mesenchymal stem cells.

Organoids derived from normal ovarian tissue

The most common source of ovarian organoids are human or mouse embryonic, fetal, and adult ovarian tissue. Organoids can be generated from fresh or frozen tissues obtained via surgical procedures, biopsies, or post-mortem ovaries. Several studies have identified two stem cell populations— very small embryonic-like stem cells (VSELs) and ovarian stem cells (OSCs)— in the adult ovaries of mice, rabbits, sheep, marmo sets, and human (9). These cells can be isolated and cultured within matrices that support 3D growth and differentiation into ovarian cell lineages. For instance, reconstruction of ovarian follicles has been achieved by seeding primary ovarian cells onto decellularized ovarian tissue (10, 11).

Fetal tissues also provide a rich source of stem cells and may recapitulate early developmental stages. However, ethical considerations and the limited tissue availability present challenges. Mizuta et al. (12) reconstructed human and monkey ovaries by culturing fetal ovarian tissue for three months, demonstrating that oogonia progressed through the first meiotic prophase, differentiated into diplotene oocytes, and formed primordial follicles. Similarly, Li et al. (13) established an ovarian organoid model derived from mouse female germline stem cells using a 3D Matrigel culture system, resulting in the production of viable offspring. These organoids also proved valuable for drug screening and toxicology studies.

Organoids derived from cancer tissue

Ovarian organoids can also derived from epithelial, serous, and other ovarian cancer cells, closely recapitulating the morphology and functionality of the primary tumor (14, 15). These organoids are often patient-specific, reflecting individual genetic mutations (3, 16-18). One key applications is drug sensitivity testing, making them valuable for personalized therapeutics. They also serve as powerful models for studying ovarian cancer biology, elucidating mechanisms of tumor formation, and evaluating potential treatment, particularly in in vitro culture systems (18).

Compared with traditional 2D cell cultures and tumor cell lines, ovarian organoids offer substantial advantages. In 2D cultures and cell lines, only a single cell type is typically used, whereas organoids can incorporate multiple cell types, including stromal, endothelial, immune, and tumor cells, thereby capturing tumor heterogeneity (19, 20). Furthermore, by employing carefully designed natural and synthetic ECMs, such as collagen, fibrin, hyaluronic acid, chitosan, and alginate, and by modifying the surrounding microenvironment, 3D culture systems can generate tumor structures that closely resemble the primary tumor (21). This allows organoids to exhibit drug and toxicant responses that more accurately reflect the physiological behavior of the original tumor.

For example, Hill et al. (22) analyzed 33 organoid cultures derived from 22 patients with high-grade serous ovarian cancer to assess defects in homologous recombination and replication fork protection, including mutations in BRCA1 or BRCA2. Their results demonstrated that combining genomic analysis with functional testing of organoids enables the identification of targetable DNA damage repair defects (22). Similarly, Chang et al. (16) successfully established primary ovarian cancer organoids from six patients, which closely resembled the original tumors in terms of morphology, histology, and genomic features. These organoids were subsequently used to evaluate drug responses for personalized therapeutic applications. Additional details on ovarian cancer organoid models can be found in several comprehensive review articles (19, 23-25).

Embryonic stem cells-derived organoids

Several studies have explored the differentiate of pluripotent stem cells—including mouse, human, and other mammalian embryonic stem cells (ESCs)— into germline and primordial germ cells (PGCs), oocytes, and somatic cells (7, 8, 26). Under appropriate culture conditions, these mixed cell populations can self-organize to form cell aggregates or ovarian organoids (27-32).

Jung et al. (27) first reported the formation of ovarian follicle-like structures in vitro using human ESCs. In their study, differentiated germ cells expressed the RNA- binding proteins DAZL and BOULE, indicating exit from pluripotency and enter into meiosis. Malik et al. (28) successfully induced goat ESCs to differentiate into PGCs and oocyte-like cells using bone morphogenetic protein-4 and trans-retinoic acid. The oocyte-like cells formed cumulusoocyte complexes (COCs) through interactions with adjacent somatic cells. These COCs exhibited a distinct zona pellucida and were positioned in close association with somatic cells. Molecular analyses confirmed the expression of oocyte-specific markers, including ZP1, ZP2, VASA, DAZL, STELLA, and PUM1. Parthenogenetic activation demonstrated that these oocyte-like cells could progress through early embryo development, reaching the morula, blastocyst, and hatched blastocyst stages (28).

The Hayashi group extended these findings by generating follicular structures using PGCs derived from ESCs and fetal ovarian somatic cells. ESCs and induced pluripotent stem cells (iPSCs) were first differentiated into primordial germ cell-like cells, which were then aggregated with female embryonic gonadal somatic cells for eight days. One month after transplantation under the ovarian bursa, the germ cell-like cells developed into germinal vesicle (GV) stage oocytes. Subsequent in vitro maturation allowed GV oocytes to reach the metaphase II stage, and fertilization with spermatozoa resulted in viable offspring. These studies highlight an alternative strategy for complete in vitro oogenesis (29, 30).

Yang et al. (31) employed iPSCs carrying a SYCP3- mKate2 knock-in reporter, generated via the CRISPR/ Cas9, to monitor meiosis during oocyte differentiation. After activating the induced PGCs/oogonia with small molecules targeting the Wnt signaling pathway, the cells were co-cultured with reconstructed human ovarian nests, effectively facilitating differentiation into primary oocytes. Similarly, Yoshino et al. (32) demonstrated well characterization PGCs derived from pluripotent stem cells, showing close similarity to embryo-derived PGCs. They developed a stepwise differentiation protocol to generate functional oocytes from pluripotent stem cellderived PGC -like cells via co-culture with embryonic ovarian somatic cells.

Induced pluripotent stem cells-derived organoids

iPSCs represent another cell valuable source for ovarian organoids. These cells are reprogrammed from embryonic or adult somatic cells and can differentiate into multiple ovarian cell types, making them excellent candidates for organoid construction (6). iPSC-derived organoids are particularly useful for studying ovarian biology and disease etiology.

Various methods exist for generating iPSCs, including the use of transcription factors such as OCT4, Sox2, and Klf4 or viral-mediated reprogramming, particularly with lentiviruses (33, 34). While these approaches are effective, potential drawbacks include gene overexpression or inactivation of unrelated genes, such as proto-oncogene and transcription factors (c-MYCNANOG, LIN28, TERT, </italic> and<italic> KLF4). Using optimized culture systems, iPSCs can form ovarian organoids suitable for investigating a range of various ovarian disorders (8, 29, 30, 34).

Wang et al. (35) proposed an effective culture method to derive oocytes in vitro from female germline stem cells. Luo et al. (36) transdifferentiated mouse spermatogonial stem cells into oocytes via transduction of H19, Stella, and Zfp57 and inactivation of Plzf genes. Following transplantation of differentiated cells into the ovaries of a premature ovarian failure mouse model, and screening for key factors within the ovarian organoids, they confirmed successful induction of functional oocytes the production of viable offspring (36).

Hikabe et al. (37) described the complete process of oogenesis from mouse pluripotent stem cells, including in vitro differentiation, growth, and maturation, generating mature oocytes from both ESCs and iPSCs derived from embryonic and adult fibroblasts. Similarly, Wang et al. (38) applied these methods to investigate the effect of MitoQ on ovarian organoid-derived oocytes. Exposure to 100 nM MitoQ enhanced follicle growth and maturation, reduced oxidative stress, and protected oocytes and granulosa cells, highlighting a potential strategy for improving oocyte quality in vitro.

Mesenchymal stem cells derived organoids

Numerous studies have demonstrated the beneficial effects of mesenchymal stem cells (MSCs) from various sources in improving ovarian function and fertility. MSCs can home to damaged ovarian tissue, proliferate in response to growth factors, and potentially differentiate into oocytes or supportive somatic cells, or alternatively, exert paracrine effects at the ovarian site (39, 40).

Mirzaeian et al. (10) reported that co-culturing oogonia with bone marrow-derived MSCs led to the formation of a greater number of follicle-like structures compared to controls. Molecular analyses confirmed differentiation into oocyte-like cells. Similarly, Li et al. (41) reconstructed an ovarian model by aggregating bone marrow-derived MSCs with neonatal mouse ovarian cells at a 1:1 ratio using phytohemagglutinin. Transplantation of these aggregates under the kidney capsule of the recipient mice revealed that MSCs did not directly form follicles but localized around growing follicles, enhancing ovarian cell survival and follicular development (41).

In a recent study from our group, human endometrial MSCs were co-cultured with mouse GV oocytes under 3D conditions to generate ovarian organoids. This study demonstrated that bidirectional inductions between oocytes and MSCs promoted organoid formation (42). Light microscopy revealed large ovoid cells with euchromatin nuclei resembling oocytes within the cell aggregates. Molecular and protein analyses confirmed the differentiation of MSCs into germ cell-like cells, indicated by alkaline phosphatase activity and expression of DDX4 and SYCP3. Furthermore, MSCs were shown to enhance oocyte survival, maturation, and development potential within the organoids (43).

Establishment of ovarian organoids

The establishment of ovarian organoids from ovarian tissue involves several critical steps (44). After careful harvesting, often requiring surgical precision to preserve tissue integrity, the ovarian tissue is dissected and enzymatically treated with agents such as collagenase, trypsin, or dispase to generate a single-cell suspension. This procedure typically combines mechanical disruption with enzymatic digestion to break down the ECM and release individual cells. Depending on the study’s objective, specific cell populations, such as oocytes, granulosa cells, or theca cells, can be isolated. The identity and purity of collected cells are commonly confirmed using techniques such as fluorescence-activated cell sorting or density gradient centrifugation.

The isolated cells are then cultured in defined media containing essential nutrients, hormones, and growth factors to support proliferation, differentiation, and organoid development. Media composition can be tailored to favor the growth of oocytes, granulosa cells, or other ovar ian cell types. Commonly used growth factors include fibroblast growth factor (FGF), epidermal growth factor (EGF), and insulin-like growth factor (IGF), which regulate cellular growth, metabolism, and survival (45).

When seeded in an appropriate matrix under optimal conditions, the cells proliferate and self-organize into 3D structures. Intercellular communication and interactions with the ECM are critical for organoid formation (23). Signaling pathways such as Wnt, Notch, and Hedgehog play essential roles in directing differentiation and spatial organization. Over days to weeks, these processes lead to the development of functional organoids that exhibit morphogenetic changes, forming distinct layers and diverse cell types through intrinsic genetic and epigenetic regulation as well as extrinsic cues from the ECM and growth factors. The presence of specific growth factors further guides differentiation, of the cells, enabling the generation of defined ovarian cell populations (46). Various scaffold-free and scaffold-based culture approaches are employed, some of which are illustrated in Figure 2.

Fig.2.

In vitro culture methods for ovarian organoids.

Scaffold-based methods

The native ovary possesses a highly dynamic and complex ECM that is influenced by both systemic and local signals. The ECM provides essential mechanical, nutritional, and inductive support for organoid formation and can modulate cellular behavior, including growth, proliferation, and differentiation. Consequently, selecting an appropriate ECM is critical for ovarian organoid culture, and choices may include natural biomaterials, hydrogels, or decellularized tissue matrices, depending on the specific experimental objectives.

Cells are seeded into an ECM that closely mimics the ovarian microenvironment to ensure effective cell–matrix interactions and transmission of appropriate biochemical and mechanical signals that promote cell growth and differentiation. Natural and synthetic materials commonly used include hyaluronic acid, gelatin, collagen, fibronectin, laminin, plasma clots, chitosan, alginate, cellulose, glycidyl methacrylate-hyaluronic acid, and polyethylene glycol (44, 47). Scaffold design aims to replicate the native tissue architecture, supporting essential cell–cell and cell–ECM interactions while providing a permissive environment for organoid development.

Hydrogels are among the most widely used biomaterials for 3D cell culture and organoid formation. They provide structural support for cell attachment and growth, facilitate the delivery of water-soluble compounds—including essential nutrients—and promote the removal of cellular waste due to their high water content (48).

Alginate is a commonly employed hydrogel in ovarian organoid research. It is a linear polysaccharide composed of repeating units of β-D-mannuronic acid and α-L-guluronic acid units, capable of forming cross-links with cationic ions such as calcium, resulting in gel formation (49). Alginate-based hydrogels have been successfully used to support the culture of ovarian tissue and follicles, demonstrating their suitability for maintaining cellular viability and promoting organoid development (43, 50, 51).

Matrigel, a basement membrane extract derived from Engelbreth–Holm–Swarm mouse sarcoma cells, closely resembles the basal lamina and is primarily composed of laminin, collagen IV, and entactin. While Matrigel is widely used for 3D ovarian cultures, it has several limitations, including its tumor origin, immunogenic variability, and batch-to-batch inconsistency. These drawbacks have prompted the development of alternative biomaterials with well-defined compositions and properties (52).

As reviewed by Dadashzadeh et al. (53) such materials not only preserve follicular architecture but also promote oocyte maturation. Oliver et al. (54) developed a modified 3D culture system in which gonadal cells were embedded in a layer of Matrigel situated between two cell-free layers of the same matrix This organized three-layer gradient system facilitated organoid formation within seven days, representing a novel approach in developmental biology and regenerative medicine for investigating mechanisms of gonadal organogenesis.

Li et al. (15) established an ovarian organoid model using mouse female germline stem cells in a 3D Matrigel- based system. The resulting organoids were capable of generating functional oocytes that produced viable offspring, demonstrating their physiological relevance. In addition, this model provided a platform for drug screening and toxicological assessment. For instance, the study showed that salinomycin adversely affected ovarian organoid formation and the germ cell population by inducing apoptosis.

Yoshino et al. (32) introduced a two-step approach for generating in vitro ovarian organoids. First, cell aggregates were formed by combining stem cell- or ovarian- derived PGCs with gonadal somatic cells or stem cell- derived follicle–like cells. These aggregates were then encapsulated in Matrigel and cultured on collagen-coated inserts. The resulting structures resembled ovarian follicles and successfully yielded oocytes.

Among the promising alternatives in the field, decellularized tissue matrices have received significant attention. These scaffolds are produced by removing cellular and genetic material from natural tissues, preserving the ECM components critical for organoid formation (55, 56). The choice of decellularization method is essential to maintain the integrity of ECM elements and bioactive molecules. Table 1 summarizes studies on the application of decellularized ovarian tissue for constructing ovarian organoids.

Table 1.

Application of decellularized ovary for ovarian organoid formation

First author(Reference)	Species / tissue source	Decellularization method	Cell sources	Key findings/results
Hassanpour et al. (57)	Human ovarian strips	Sodium lauryl ester sulfate and DNase I	Primary ovarian cells	Formation of primordial or primary follicle-like structures within scaffolds after transplantation.
Mirzaeian et al. (10)	Human ovarian fragments	NaOH solution and RNase/DNase	Oogonial, peritoneal, and bone marrow mesenchymal stem cells	Differentiation into oocyte-, germ-, and granulosa-like cells; estradiol production.
Amjadi et al. (58)	Bovine ovarian tissue	Chemical cocktail	Cumulus cells and human endothelial cells	Estrogen production.
Wu et al. (59)	Whole porcine ovaries	Sodium deoxycholate, Triton X-100 and RNase/DNase	Mouse granulosa cells and follicles	Artificial ovaries formed but did not restore endocrine function.
Sistani et al. (60)	Human ovarian fragments	Freezing/thawing cycles; 1% Triton X-100 and SDS-based solutions	Mouse fetal ovarian cells	Follicle structure formation confirmed; expression of oocyte- and follicular-specific genes observed.
Alaee et al. (61)	Rat ovaries	Sodium lauryl ester sulfate and DNase I	Mouse preantral follicles	High follicular and oocyte maturation rates; elevated hormone production.
Pennarossa et al. (13)	Porcine whole ovaries	Sodium dodecyl sulfate, Triton X-100 and deoxycholate	Porcine ovarian cells and human dermal fibroblasts	Both porcine and human cells repopulated and adhered to scaffolds; ovary-specific gene expression confirmed.
Zheng et al. (12)	Swine ovarian tissue	Production of bioink from decellularized matrix	Primary ovarian cells	Enhanced neoangiogenesis, cell proliferation, and survival; hormone production achieved via 3D bioprinting.
Park et al. (62)	Porcine ovarian cortex	Trypsin-EDTA, Tris(hydroxymethyl)aminomethane and sodium deoxycholate	Mouse preantral follicles	High rates of oocyte and follicular maturation.
Laronda et al. (63)	Bovine and human ovaries	0.1% Sodium dodecyl sulfate	Mouse primary ovarian cells	In vitro estradiol production; initiation of puberty observed after transplantation in mice.

Recent advancements have focused on employing mechanical, chemical methods, or combined approaches to decellularization in order to minimize damages to the 3D structure, ultrastructure, and mechanical properties of the ECM. These methods aim to preserve the connective components and adhesion proteins, ensuring that the resulting scaffold closely mimics the native tissue architecture (64). Such preservation not only facilitates cell attachment, infiltration, and proliferation but also promotes differentiation toward organoid formation, providing a distinct advantage over conventional 2D cultures (65).

Decellularized tissue matrices can be utilized in three forms: as whole ovaries, tissue fragments, or hydrogels derived from decellularized tissue. This approach is particularly attractive because it closely mimics the native ovarian composition and retains essential components required for oocyte and follicle growth, maturation, and development, facilitating effective cell–scaffold interactions. Nevertheless, challenges remain, including the maintenance of tissue architecture and the preservation of critical molecules necessary for cell adhesion, proliferation, and differentiation.

During decellularization, the use of detergents or mechanical and chemical methods can alter or damage the tissue composition, potentially affecting its interaction with the cells. However, several studies have focused on optimizing decellularization protocols to minimize such effects and better preserve the native matrix composition (10, 46, 57, 59, 60, 66, 67).

Hassanpour et al. (61) decellularized human ovarian scaffolds using various methods and subsequently seeded primary ovarian cells onto the resulting constructs. These scaffolds were then transplanted under the kidney capsule, and in vivo analysis confirmed the presence of the primordial or primary follicle-like structures within the transplanted scaffolds. Similarly, Mirzaeian et al. (10) demonstrated that decellularized human ovarian matrices could support oogenesis and the reconstruct follicular structures by seeding oogonial, peritoneal, and bone marrow-derived mesenchymal stem cells. Another study showed that the function of cumulus and endothelial cells could be restored following transplantation of a decellularized bovine ovarian niche seeded with cumulus cells and human endothelial cells into a rat subcutaneous site (58). Wu et al. (59) developed an effective ovarian tissue decellularization protocol, followed by recellularized with granulosa cells and follicles. After two weeks of in vitro culture, granulosa cells attached to the scaffold and proliferated; however, despite follicular growth, the artificial ovary did not exhibit endocrine function after subcutaneous transplantation.

Recently, our group investigated the supportive role of the decellularized human ovarian tissue in promoting the homing of mouse fetal ovarian cells and the reconstruction of follicular-like structures. Our results demonstrated that this scaffold effectively supports follicular formation and provides a reliable in vitro model for ovarian organoid generation (60). Similarly, Zheng et al. (11), reconstructed ovarian follicles using bioink derived from decellularized swine ovary, which was combined with primary ovarian cells through 3D bioprinting. Their study highlighted this approach as a promising alternative for ovarian failure treatment.

Pennarossa et al. (66) decellularized entire porcine ovaries and subsequently recellularized the matrices with porcine ovarian cells or human dermal fibroblasts. Their result demonstrated that this approach produced an optimal 3D structure for ex vivo ovarian cell culture and facilitated the formation of ovarian organoids. Similarly, Alaee et al. (61) compared follicular development and oocyte maturation in decellularized ovarian matrices (3D) with conventional 2D culture systems. They found that follicular development, hormone production, and oocyte maturation rates were higher in the 3D cultures, closely resembling the natural ovary.

Non-adhesive surfaces are engineered to minimize unintended adhesion of cells, proteins, or other materials, making them valuable in cell culture, tissue engineering, and biomedical research. These surfaces are particularly useful for culturing cell types that require minimal attachment or for promoting cell aggregation and spheroid formation. They can be modified with hydrophilic polymers, such as polyethylene glycol, to reduce protein adsorption and cell attachment. Additionally, surface topography can be engineered at the micro- or nanoscale for example, enable cells to cluster and self-organize in three dimensions, enhancing the physiological relevance of in vitro models. Consequently, non-adhesive surfaces facilitate organoid formation by allowing progenitor cells to aggregate and self-organize into structures that closely mimic native organs (62, 63).

Organ-on-a-chip technology involves the development of micro-engineered devices that incorporate living cells to replicate the mechanical and biochemical functions of specific human organs. These systems aim to reproduce complex tissue architecture and physiological responses, providing a relevant platform for studying biological processes. Typically constructed from biocompatible materi als, such as polydimethylsiloxane, these devices feature microchannels and chambers that enable precise control of flow rates, shear stress, and nutrient delivery, mimicking blood flow or interstitial fluid movement. The device facilitates replication of cellular organization and structural features of the target organ and often incorporating multiple cell types to recreate the organ’s microenvironment and intercellular interactions. Organ-on-a-chip systems can also be equipped with sensors and imaging tools to monitor cellular behavior, biochemical outputs, and physiological responses in real time, allowing high sensitivity assessment drug effects, toxicity, and disease modeling (68).

Scaffold free methods

Scaffold-free culture systems, such as hanging drops and spinner flasks, enable the grow of cells and tissues in three dimensions without the need for an external supporting scaffold (42, 44). These methods promote natural cell behavior, and each approach offers distinct advantages depending on the specific requirements of the study. In the hanging drop method, a small volume of cell suspension is plated on the underside of a culture dish lid or a specialized plate. Gravity causes the cells to aggregate at the drop, facilitating cell-cell interactions and leading to the formation of spheroids or tissue-like structures.

Recently, we developed a novel strategy for ovarian organoid formation using the hanging drop method with human endometrial mesenchymal stem cells, compared with a 3D culture system employing sodium alginate hydrogel. In both approaches, differentiation was induced through co-culture with mouse GV oocytes. Our results demonstrated that the hanging drop method was more effective in generation organoid structures compared with the sodium alginate hydrogel system (42).

The spinner flask is a well-established method for cultivating cells in a controlled 3D environment. The dynamic conditions generated by the spinning mechanism enhance cell-cell interactions and better mimic the natural tissue environments, which is crucial for developing functional tissue. This approach maintains a homogenous cell suspension while improving nutrient and oxygen exchange. Optimization, including adjustments to flask size, oxygen concentration, and nutrient supply—such as those implemented in bioreactor systems —further enhance its performance (69). Although spinner flasks have been widely used for generating various organoid types, their application in ovarian organoid culture has been limited (69).

Applications of ovarian organoids

Ovarian organoids derived from normal or cancerous tissues have a wide range of potential applications, including basic ovarian research, fertility studies and treatment, drug testing and screening, cancer biology and therapeutics, disease and cancer modeling, endocrine function analysis, personalized medicine, and the investigation of host–pathogen interactions. Selected applications are illustrated in Figure 3.

Fig.3.

Potential applications of ovarian organoids.

Cancer biology, treatments and modeling

Ovarian cancer is among the most common gynecological malignancies, with over 70% of cases diagnosed at advanced stages. Conventional treatments for advanced ovarian cancer are often limited in effective, resulting in poor prognoses and low survival rates (70). Consequently, alongside emerging targeted therapies and immunotherapies, tumor markers are increasingly employed for early disease detection. A deeper understanding of ovarian cancer biology is essential to improve patient outcomes. In this context, ovarian organoids have emerged as a powerful tool in cancer research (70, 71). These organoids preserve the genetic and phenotypic characteristics of the primary tumors, enabling insights into cancer etiology and facilitating the identification of effective therapeutic agents. Their stability during prolonged culture and close resemblance to primary cancer cells make organoids particularly valuable for drug testing, screening, and resistance studies, offering advantages over conventional 2D cell culture models (71-73).

Previous studies have commonly relied on cancer cell lines, spheroids, and animal models (73, 74). To date, over 50 ovarian cancer cell lines have been employed in research (75); however, these approaches often result in the loss of stromal cells and ECM during successive passages, rendering them substantially different from the original tumor tissue. Moreover, genetic alterations can accumulate during culture, producing gene expression profile that diverge from those of the primary cancer. As a result, these models lack the heterogeneity of primary tumors and are limited in their suitability for drug testing and translational cancer research (75).

In animal studies, tumor tissues are commonly xenotransplanted into sites such as such as subcutaneous tissue, the primary tumor location, or the peritoneal cavity (76, 77). Several research groups have successfully developed ovarian cancer models using this approach. Although these models are valuable for drug studies, they are limited by their time- intensive nature and high costs (76, 77).

Organoid technology has emerged as a transformative tool in cancer research, enabling the study of cancer etiology, including the roles of pathogens and oncogenic genes. Furthermore, pharmacological interventions can be precisely evaluated by assessing the effects of therapeutic agents on organoids derived from patient-specific or cancer cell lines, providing a more physiologically relevant platform for preclinical studies (78, 79).

Kopper et al. (80) developed a robust approach for the long-term expansion of ovarian cancer organoids, establishing 56 organoid lines from 32 patients and encompassing a broad spectrum of ovarian cancer subtypes. Characterization of these organoids revealed both intra- and interpatient heterogeneity, reflecting the complexity of primary tumors. The organoids demonstrated suitability for drug screening, including responses to platinum- based chemotherapy, underscoring their potential in both research and personalized medicine. Several chemotherapeutic agents—such as cisplatin, paclitaxel, bevacizumab, and olaparib—were tested on these organoids, revealing variable yet high responsiveness. These findings support the use of ovarian cancer organoids to these drugs. Due to the variable responses observed, they suggested that these chemotherapeutic agents could be tailored for use in personalized as a platform to guide individualized treatment strategies.

Zhang et al. (78) generated organoids from fallopian tube epithelium and ovarian surface epithelium, each carrying identical oncogenic alterations, and observed differential chemosensitivity between the models. These findings suggest that the cell of origin may influence therapeutic responses.

Kwong et al. (81) established an in vitro organoid model from normal human ovarian surface epithelial cells. The resulting spheroid resembled epithelial inclusion cysts in the ovarian cortex, providing evidence to support the link between chronic inflammation and ovarian cancer development and offerings insight into the disease’s etiology.

Innovative organoid models allow the incorporation of cancer cells with immune cells, enabling the study of inter actions within the tumor microenvironment and supporting the development of novel strategies in immune-oncology (82). Recent studies have focused on co-culturing ovarian cancer organoids with autologous immune cells to evalu ate therapeutic responses. These approaches aim to activate natural killer cells and CD⁺⁸ T cells, promoting cytotoxic phenotype and enhancing anti-tumor efficacy (8, 82).

Drug assessment, screening, and toxicity

Organoids provide a robust platform for high-throughput drug screening, enabling the evaluation of both the efficacy and toxicity of pharmacological agents in a physiologically relevant context (16). Unlike traditional 2D cultures that rely on a single cell type, organoid models incorporate multiple cell types, creating structures that closely mimic native tissue and produce drug responses more reflective of in vivo conditions. Patient-specific organoids derived from autologous cells further facilitate the development of personalized therapies. Additional advantages of organoid- based drug screening include reduced testing time and lower associated costs (83, 84). Organoid systems are also suitable for preclinical toxicity testing, enhancing the safety assessment of new compounds prior to clinical trials (4). For example, Chitrangi et al. (83) demonstrated that ovarian and breast cancer organoids exhibited heterogeneity in drug sensitivity and resistance and sensitivity, highlighting their utility for individualized therapy design.

Basic ovarian research

Ovarian organoids provide a powerful platform for investigating the mechanisms underlying folliculogenesis and oocyte maturation. This system enables detailed assessment of interactions between somatic cells, including follicular and theca cells, and oocytes, thereby enhancing our understanding of the cellular crosstalk and regulatory factors that govern follicular development.

Ovarian organoids can recapitulate the native ovarian microenvironment, enabling the study of oocyte development and fertilization. This model supports investigation from the early stages of follicle growth through to oocyte maturation. It is particularly valuable for elucidating the differentiation processes from stem cells to PGCs, germline cells, and ultimately, mature oocytes. The system also facilitates the study of interactions between germ cells and somatic cells, with signaling pathways such as BMP, WNT, and retinoic acid playing critical roles in germ cell development (6, 7).

Through the introduction of genetic variations or targeted gene knockouts, researchers can delineate the specific roles of individual genes in oocyte development and assess their impact on oocyte quality under abnormal conditions. Additionally, the expression of key regulatory genes involved in germ cell specification and oocyte maturation can be monitored. These studies provide critical insights into how gene activation or repression directs germ cell fate.

In addition, ovarian organoid derived from genetically modified stem cells provide a platform to investigate the effects of specific gene mutations on germ cell development. This approach is particularly valuable for elucidating the genetic basis of reproductive disorders and infertility, as it enables direct correlation between genetic alterations and defects in germline cell formation and oocyte quality (8).

The transition from stem cells to germ cells involves extensive epigenetic remodeling. Ovarian organoids provide a valuable system to investigate these epigenetic modifications and their impact on gene expression and cellular differentiation, offering insights into the stability and inheritance of epigenetic marks in germline cells (7, 8).

Fertility research and treatment

In vitro ovarian organoids offer a powerful platform for advancing fertility research and assisted reproductive techniques. These organoids enable the evaluation of hormonal regulation, including the roles of gonadotropins in ovarian function, facilitating the optimization of hormone treatments for estrus synchronization and overall reproductive efficiency.

Cryopreservation has long been employed in assisted reproductive technologies, and ongoing improvements continue to enhance fertility preservation for various individuals, including cancer patients. In this context, ovarian tissue, oocytes, or embryos can be cryopreserved prior to gonadotoxic treatments such radiotherapy or chemotherapy, providing an effective strategy for fertility preservation (85).

Despite these advances, limitations in obtaining human samples—both in quantity and due to ethical considerations—highlight the need for alternative models. Ovarian organoids represent a promising solution, serving as a platform to assess, optimize, and refine cryopreservation techniques and related interventions.

Understanding the biochemical and physiological responses of ovarian organoids to cryopreservation can improve the efficiency of oocyte and embryo freezing and thawing, thereby enhancing assisted reproductive technologies such as in vitro fertilization.

Moreover, ovarian organoids provide a valuable platform for evaluating drug toxicity during oocyte development and maturation and for the development of novel therapeutics in infertility treatment and fertility preservation.

Assisted reproduction in farm animals

In vitro ovarian organoid culture can be applied to promote follicle growth and development across various mammalian species, such as cattle, sheep, and pigs. This approach enables detailed investigation of folliculogenesis, oocyte maturation and fertility-related issues, such as irregular estrous cycles or low conception rates, providing insights into the underlying physiological and molecular mechanisms and facilitating the identification of potential interventions (26).

Additionally, ovarian organoids offer a platform to assess the effects of dietary regimens and management practices on ovarian function, ovulation rates, and overall reproductive performance in farm animals.

Ovarian organoids can be employed to evaluate the effects of environmental toxins or feed additives on ovarian health and reproductive outcomes in livestock, providing guidance for safer agricultural practices. This knowledge can inform breeding programs aimed at enhancing reproductive traits while ensuring animal welfare.

Furthermore, ovarian organoid technology holds significant potential for the conservation of rare and endangered species. By enabling the generation of large numbers of female gametes or oocytes, this approach can accelerate fertility processes, increase birth rates, and offer a costeffective strategy for species preservation.

Personalized medicine

Ovarian organoids derived from individual patients enable the development of personalized treatment strategies, representing a major advance in precision medicine (5, 19, 24). In oncology, patient- specific organoids can be generated from tumor samples to test chemotherapy agents and targeted therapies. By evaluating the responses of these organoids, clinicians can better predict the efficacy of treatments for each patient.

Given the genetic heterogeneity of ovarian cancer, organoid technology allows the study of diverse cell populations within a tumor, facilitating the identification of mutations and biomarkers that may influence therapeutic decisions. Organoids also provide a platform to examine the role of the tumor microenvironment in treatment responses, which is crucial for tailoring therapies.

Furthermore, organoids can be employed to investigate mechanisms of drug resistance, guiding the design of combination therapies to overcome these barriers. Integration with advanced imaging techniques allows precise monitoring of treatment responses, further enhancing the capacity for individualized therapy planning.

Ovarian endocrine function study

Ovarian organoids provide a powerful platform to investigate hormonal signaling pathways and their impact on reproductive function and disorders. These organoids can replicate key aspects of ovarian endocrine activity, including hormone synthesis and regulation, allowing controlled studies of complex hormonal interactions.

By monitoring hormone secretion patterns, researchers can assess how endocrine factors influence the responses to therapeutic agents, offering insights for personalized treatment strategies. Organoids can also model physiological fluctuations in hormone levels across different menstrual cycle phases, which is crucial for understanding hormonal in fluences on ovarian cancer progression and therapy efficacy.

Additionally, ovarian organoids can be used to study conditions such as polycystic ovary syndrome or ovarian dysfunction, providing insights into hormonal imbalances and their effects on ovarian health (86). They enable the evaluation of follicular development and oocyte maturation in response to endocrine signals, enhancing our understanding of fertility and reproductive physiology. Studies of ovarian organoids can also elucidate how hormones, such as estrogen and progesterone, affect tumor growth, differentiation, and treatment responses, facilitating the development of targeted therapies that account for individual endocrine profiles (26).

Pathogen interaction study

Ovarian organoids provide a physiologically relevant 3D model to investigate interactions between pathogens, such as viruses and bacteria, and ovarian epithelial cells. This system enables detailed studies of infection mechanisms and pathogen replication within ovarian tissues. Organoids have been used to examine viral entry and replication for pathogens including human immunodeficiency virus (HIV), hepatitis B and C, human papillomavirus (HPV), and Zika virus, offering insights into viral pathogenesis (87).

Co-culture of ovarian organoids with immune cells allows researchers to assess host immune responses to infection and their impact on ovarian function and tissue integrity. This approach facilitates the study of pathogen- induce inflammation within the ovarian microenvironment and its potential role in ovarian dysfunction or disease. Furthermore, ovarian organoids provide a platform evaluating the efficacy of antiviral or antibacterial interventions, supporting the development of targeted therapies that preserve ovarian function. Investigating pathogen interactions with ovarian tissues also informs potential risks to fertility and reproductive health, guiding strategies for protective and therapeutic strategies in individuals.

Extracellular vesicles and ovarian organoid

Extracellular vesicles (EVs) are membrane-bound nanoparticles secreted by cells that carry bioactive molecules, including proteins, lipids, and nucleic, facilitating intercellular communication. EVs are commonly classified based on their size into microvesicles (100-1000 nm), exosomes (30-150 nm), and apoptotic bodies (50-5000 nm) based on their size (88).

EVs secreted by the reproductive system, such as those present in follicular fluid, play critical roles in oocyte maturation and development. Similarly, EVs released in the fallopian tubes contribute to sperm viability, motility, and functional maturation. In vitro studies have further demonstrated that EVs derived from cultured cells can positively influence oocyte maturation as well as sperm quality and functionality (89, 90).

Recent research has increasingly focused on EVs released from organoids derived from both normal and tumor cells. These studies reveal that EVs from organoids closely resemble those from native tissues, a similarity not observed in conventional two-dimensional cultures. This highlights the structural and functional fidelity of organoids, emphasizing their utility for studying tissue biology and reproductive processes in a physiologically relevant context (89, 90).

Organoid culture also enables the study of EVs released from specific cell types, a level of specificity not achievable in vivo. Although numerous studies have examined EVs from various organoids, investigations focusing on reproductive system-derived organoids remain limited. Most reports have addressed organoids from the endometrium and fallopian tubes, with ovarian organoids yet to be explored. Nevertheless, research on EVs holds considerable potential for diagnostic and therapeutic applications, particularly in infertility treatments, by enhancing oocyte and sperm quality and providing insights into ovarian physiology and pathology (89, 90).

The successful establishment of ovarian organoids is typically confirmed through morphological assessment, immunohistochemistry, and molecular analyses (Fig .4).

Fig.4.

Characterization of ovarian organoids. A. Organoids were assessed morphologically by light microscopy, B. For protein expression by immunohisto chemistry, and C. For gene expression by molecular analysis. D. Functional activity was evaluated by analyzing secreted factors and extracellular vesicles (EVs).

Challenges in ovarian organoid research

Despite significant advances in the establishment and application of ovarian organoids, several challenges remain. The efficiency of organoid generation can be inconsistent, influenced by tissue techniques, and donor variability. Different ovarian cell sources contribute to variability in organoid morphology and function. Maintaining long-term viability and functionality of human ovarian cells in vitro is particularly challenging, as granulosa cells, stromal cells, and oocytes, often exhibit limited lifespans and can lose specialized functions over time. This limitation impacts studies requiring prolonged culture, such as chronic drug testing or longitudinal analyses of ovarian tissue development.

Although organoids can proliferate and expand, their longterm maintenance is difficult, often resulting in cellular senescence or loss of functional characteristics. Strategies to address these challenges include standardizing cell sources and culture conditions, employing optimized media formulations enriched with growth factors, hormones, and antioxidants, and using microfluidic devices or bioreactors to simulate blood flow, nutrient exchange, and waste removal, thereby enhancing longevity and functionality.

Reproducing the intricate architecture and multicellular interactions of the ovary remains another major challenge. The ovary comprising follicles at various developmental stages, stromal cells, blood vessels, nerve fibers, and ECM components. Accurately mimicking this complexity is essential for realistic modeling of ovarian functions, including the stromal-epithelial interactions and hormone signaling. Variability in human tissue samples due to genetic background, age, or health status can further introduce inconsistent in organoid characteristics, affecting reproducibility and comparability across studies.

For tumor-derived organoids, additional considerations include faithfully recapitulating the tumor microenvironment to ensure accurate modeling of cancer biology. Collectively, standardizing of protocols for organoid generation, culture, and characterization is critical to improve reproducibility, reliability, and translational potential of ovarian organoid research (84).

Genetic heterogeneity and CRISPR-Cas9 applications in cancer organoids

Cancer organoids exhibit genetic heterogeneity, reflecting the diverse genetic mutations within and between native tumors. This heterogeneity complicates the development of effective treatments, as different regions of a tumor may respond variably to therapy. Advanced gene-editing strategies, such as CRISPR-Cas9, provide powerful tools to mitigate the confounding effects of genetic heterogeneity in cancer organoids (90). By enabling precise genetic modifications, CRISPR-Cas9 allows researchers to standardize specific mutations and investigate their functional consequences, thereby enhancing our understanding of tumor biology and supporting the development of personalized therapies or gene expressions. CRISPR-Cas9 facilitates several applications in cancer organoid research. First, it allows the editing of specific genes across multiple organoid lines, creating uniform models that isolate the effects of individual genetic alterations and reduce variability caused by heterogeneity. For instance, introducing a defined mutation in genes such as KRAS or TP53 across organoid lines enables controlled comparisons of treatment responses. Second, CRISPR can be employed to introduce or correct mutations in oncogenes or tumor suppressor genes, allowing the functional analysis of these mutations in tumor behavior and drug sensitivity. Third, the generation isogenic organoid lines—genetically identical except for targeted mutations—provides a controlled platform to dissect the impact of single or combinatorial genetic changes on heterogeneity and therapeutic response (90).

Collectively, these applications underscore the potential of CRISPR-Cas9 to standardize cancer organoid models, reduce variability, and provide mechanistic insights into the genetic determinants of tumor behavior and treatment efficacy.

Challenges in organoid maturation, drug screening, and human tissue use

Achieving fully mature ovarian organoids capable of supporting complete oocyte growth and development remains a significant challenge and a central focus of ongoing research. In the context of drug screening, careful consideration must be given to the design of the drug construct, its assessment, and the validation of findings, including correlation with clinical outcomes (83, 84).

Accessing high-quality human ovarian tissues, particularly fetal samples, raises important ethical considerations and requires appropriate consent and adherence to regulatory guidelines. Additionally, cost-effectiveness and re imbursement remain crucial factors in the application of organoid technologies in both research and clinical settings (83).

Ethical consideration

The establishment and use of ovarian organoids raise several important ethical considerations. Donors of ovarian tissue or cells must provide informed consent, fully understanding the purposes of the research, potential applications of the organoids, and any genetic modifications involved. The use of technologies such as CRISPR-Cas9 introduces additional ethical concerns, particularly regarding germline editing. While organoids are primarily used for in vitro studies, their potential application in reproductive contexts raises the risk of creating heritable genetic changes.

Efforts to restore or generate reproductive tissue using ovarian organoids also prompt ethical questions related to cloning, artificial reproduction, and germline modifications. The creating functional ovaries or gametes raises complex debates regarding reproductive rights, identity, and parenthood, as well as regulatory and societal implications (3, 24, 83).

Organoids derived from embryonic or fetal tissues introduce further ethical considerations concerning tissue source, consent, and moral status, and are subject to strict regulatory oversight in many countries. iPSC-derived ovarian cells offer an ethically acceptable alternative when access to primary fetal tissue access is limited (3).

Overall, establishing clear guidelines and regulations for the use of ovarian organoids—particularly regarding genetic modifications and reproductive applications—is essential. While these models provide unprecedented opportunities to advance understanding of female reproductive health and develop novel therapies, their use must adhere to ethical standards, respect donor rights, and consider long-term societal impacts (83).

Future outlook of ovarian organoid research: from in novation to clinical translation

The future of ovarian organoid research is highly promising, fueled by rapid advancements in bioengineering, molecular characterization, and personalized medicine. In the short-term, efforts will focus on refining organoid cultivation techniques, standardizing protocols to ensure reproducibility, and integrating advanced bioengineering technologies—such as 3D bioprinting and microfluidic systems—to generate more physiologically relevant and complex ovarian models. These approaches will facilitate detailed functional studies, including hormone production, folliculogenesis, and tissue responses to environmental stressors.

Long-term objectives aim to establish ovarian organoids as robust platforms for disease modeling, drug screening, and regenerative therapies. The development of genetically and functionally matured organoids capable of supporting oocyte growth has the potential to transform fertility preservation and the treatment of endocrine disorders. Furthermore, these models may provide critical insights into reproductive health disparities, the impact of environmental toxins on ovarian function, and strategies for personalized medicine.

Key challenges and potential solutions

Limited access to human samples

The availability of high-quality human ovarian tissue is constrained by ethical, legal, and logistical considerations, limiting both the scale and diversity of organoid models. Alternative strategies include the use of iPSCs-derived ovarian cells and synthetic scaffolds that recapitulate ovarian tissue, reducing reliance on scarce human samples. Collaborations with biobanks and the establishment ethical tissue-sharing frameworks can further expand access.

Model maturation and functional fidelity

Achieving fully functional, mature ovarian tissue, particularly capable of supporting oocyte development, remains a significant challenge. Advanced bioengineering approaches—such as vascularization scaffolds, perfusion systems, and precise delivery of growth factor s—enhance organoid maturation and improve functional fidelity.

Reproducibility and standardization

Variability in organoid formation and characterization can hinder translation and clinical implementation. Establishing industry-wide standardized protocols, defining quality control metrics, and reaching consensus on characterization criteria— supported by multi-center validation studies—are essential to improve reproducibility.

Ethical and regulatory hurdles for clinical use

Genetic modifications, reproductive applications, and stem cell sourcing raise important ethical considerations. Early engagement with regulatory authorities, adherence to transparent research practices, and development of comprehensive ethical guidelines emphasizing safety, informed consent, and societal implications are crucial.

Long-term stability and safety for reproductive appli cations

Ensuring the long-term safety and genetic stability of organoid-derived tissues, particularly for reproductive purposes, is critical. Longitudinal studies to monitor genetic integrity, incorporate of safety mechanisms (e.g., kill switches), and strict regulatory oversight are recommended to mitigate risks.

Conclusion

Despite existing limitations and challenges, ongoing research and technological advancements in ovarian organoid development hold significant promise for advancing reproductive health and disease management. Numerous issues related to their applications in clinical settings— particularly in reproductive and regenerative medicine— require further investigation. A major challenge is the limited availability of high-quality human samples due to ethical, legal and logistical considerations. The development of alternative models that closely replicate human ovarian tissue or the creation of artificial systems may help address these limitations and expand the translational potential of ovarian organoid research.

Author’s Contributions.

M.S.; Supervised the study and Prepared the manuscript. S.S.M.; Contributed to writing and English editing of the manuscript. Both of the authors read and approved the final manuscript.