GyneCult™ Fallopian Tube Organoid Medium

Serum-free and sex steroid-modular kit for the expansion and differentiation of human fallopian tube organoids

GyneCult™ Fallopian Tube Organoid Medium

Serum-free and sex steroid-modular kit for the expansion and differentiation of human fallopian tube organoids

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Serum-free and sex steroid-modular kit for the expansion and differentiation of human fallopian tube organoids
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Product Advantages


  • Generate functional fallopian tube organoids in 10 days with a convenient and complete workflow

  • Control hormone levels to answer your specific research questions with a sex steroid-modular kit

  • Recapitulate the fallopian tube in vivo morphology with ciliated and secretory cell representation

  • Establish patient-derived fallopian tube organoids with high culture initiation and success rates

  • Minimize inter- and intra-donor variability with an optimized, serum-free medium system for reproducible experiments

What's Included

  • GyneCult™ Fallopian Tube Organoid Basal Medium, 90 mL
  • GyneCult™ Fallopian Tube Organoid Medium 5000X Supplement, 1.5 mL
  • GyneCult™ Fallopian Tube Organoid Medium 10X Supplement, 10 mL
  • GyneCult™ Fallopian Tube Organoid Medium 100X Supplement, 1.0 mL
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.

Overview

Generate mature fallopian tube (FT) organoids that exhibit a representative combination of ciliated and secretory cells as they appear in vivo. GyneCult™ Fallopian Tube Organoid Medium (FTOM) outperforms do-it-yourself (DIY) formulations by supporting the superior growth of functional FT organoid cultures with physiological proportions of relevant cell types.


Organoids generated with GyneCult™ FTOM provide a robust and versatile system for female reproductive biology research. With this optimized workflow, you can cut down on time and labor by seeding directly into 3D culture, enabling the growth of passageable FT organoids by Day 10. GyneCult™ FTOM is a serum-free and sex steroid-modular kit, by omitting the addition of the 5000X supplement, you can adjust hormone levels as needed while delivering consistent results for your specific applications.


Applications of FT organoid cultures include the study of fertility, hormone dynamics, menstrual and post-ovulation cycles, and ovarian cancer. With GyneCult™ FTOM, a subset of high-grade serous ovarian cancer (HGSOC) organoids can be cultured for use in ovarian cancer research, such as evaluating the tumor initiation capacity of FT epithelial cells or screening of chemotherapy drugs for the treatment of ovarian cancer.
Subtype
Specialized Media
Cell Type
Epithelial Cells
Application
Cell Culture, Differentiation, Expansion, Organoid Culture
Brand
GyneCult
Area of Interest
Cancer, Disease Modeling, Drug Discovery and Toxicity Testing, Epithelial Cell Biology, Organoids
Formulation Category
Serum-Free

Data Figures

Generation of Fallopian Tube Organoids Using GyneCult™ Fallopian Tube Organoid Medium

Figure 1. A Subset of High-Grade Serous Ovarian Cancer Organoids Can Be Cultured in GyneCult™ FTOM and Recapitulate Parental Tumor Biology in 3D

(A) Workflow showing the generation of fallopian tube (FT) organoids. Freshly isolated or cryopreserved human FT cells are embedded at 4000 cells per 20 μL Matrigel® dome into 24-well non-tissue culture plates. Once the domes solidify, 500 μL of GyneCult™ Fallopian Tube Organoid Medium (FTOM) is added. By Day 10, cultures contain functional cells with beating cilia and can be passaged for further culture or cryopreservation. (B) Representative images of FT organoid morphologies throughout the GyneCult™ FTOM workflow. Scale bar = 200 μm.

Differentiated fallopian tube organoids display consistent and standard morphology across three independent donors and is maintained from passage to passage.

Figure 2. GyneCult™ Fallopian Tube Organoid Medium Generates Consistent and Standardized Organoid Cultures

Dissociated human FT cells were seeded and cultured following the recommended protocol at 4000 cells per dome. Organoid cultures from passage 0 (A, D, G), passage 3 (B, E, H), and passage 5 (C, F, I), at 10 - 14 days post-seeding, are shown for three independent donors. Within each dome, FT organoids were typically cystic, with a prominent lumen and between 50 - 500 µm in diameter. Size distribution and diverse morphology within each culture were maintained from passage to passage. GyneCult™ FTOM supports human FT cultures with high consistency between donors and low passage-to-passage variability, while also preserving the morphological heterogeneity within each culture. Scale bar = 200 µm.

GyneCult™ Fallopian Tube Organoid Medium Generates Robust Cultures Without the Need for Additional Purification Steps

Figure 3. GyneCult™ Fallopian Tube Organoid Medium Generates Robust Cultures Without the Need for Additional Purification Steps

GyneCult™ FTOM supports passaging FT organoids as single cells or clumps and demonstrates a high establishment rate in the donors tested (n = 8), even with cultures that show poor initiation capabilities (e.g. Donor 3, 5 and 7). (A) FT organoids imaged at passage 3 (Donor 1), passage 4 (Donor 2), and passage 5 (Donors 3 - 8). Scale bar = 500 µm. (B) Representative flow cytometric dot plot of a dissociated donor sample stained with EpCAM and CD45 antibodies. Stromal or immune cell contamination could be present and is selected out with subsequent passages in GyneCult™ FTOM without additional purification steps. (C) Box plot showing the organoid-forming efficiency (OFE; the fraction of single cells that form organoids in culture) of dissociated FT cells is 16 ± 8% when normalized to the EpCAM+ population (mean ± SD, n = 10, includes 2 replicates).

 GyneCult™ FTOM Supports Proliferation From Fresh or Cryopreserved Samples

Figure 4. GyneCult™ FTOM Supports Proliferation from Fresh or Cryopreserved Samples

GyneCult™ FTOM supports robust organoid formation from cryopreserved or freshly isolated FT cells. (A) Organoid morphology after establishing cultures from freshly dissociated single cells or revived after cryopreservation. Shown are organoids at the end of primary culture (top) and after passage 3 (bottom). Scale bar = 500 µm. (B) Throughout the 10 - 13 day primary passage, the proliferative capacity of fresh and cryopreserved cultures is expressed in doublings per day, with no significant difference. (C) Additionally, cultured FT cells retain their organoid forming efficiency (OFE) after thawing, fluctuating between 5 - 10%, as seen in the box and whiskers plot for at least five passages. (D) Graph showing individual cryopreserved FT donors (n = 7), the samples were able to undergo 20 cumulative doublings within 5 passages, further demonstrating the ability to achieve strong organoid cultures from cryopreserved samples.

Differentiated Fallopian Tube Organoids Exhibit a Representative Mixture of Ciliated and Secretory Cell Types

Figure 5. Differentiated Fallopian Tube Organoids Exhibit a Representative Mixture of Ciliated and Secretory Cell Types

Dissociated primary FT cells from three individual donors were cultured in GyneCult™ FTOM and cultured for five passages. The resulting organoids were representative of in vivo FT biology, containing a mixture of ciliated and secretory cells as confirmed by marker antigen expression. (A-C) Cultures were stained for the secretory transcription factor PAX8 (green), and a functional component in ciliated cells—acetylated tubulin (red). (D-F) FT organoids were also stained for the secretory glycoprotein OVGP1 (green) and ciliated cell transcription factor FOXJ1 (red). These results demonstrate the efficient generation of multilineage organoids across different donors, mimicking the in vivo biology. Scale bar = 50 µm.

GyneCult™ FTOM Supports Robust Growth and High Fidelity Modeling Compared to DIY Medium

Figure 6. GyneCult™ FTOM Supports Robust Growth and High Fidelity Modeling Compared to DIY Medium

FT cells from the same donor were seeded and cultured side-by-side using (A) DIY medium and (D) GyneCult™ FTOM. The organoids were stained with IHC for (B, E) secretory transcription factor, PAX8, and (C, F) ciliated cell transcription factor, FOXJ1, both shown in brown. Blue stains indicate nuclei and the absence of nuclear expression of the target antigen. The DIY-derived organoids were dominated by (B) PAX8+ and (C) lacked FOXJ1+ markers, suggesting that these organoids consist mainly of secretory cells. (E, F) GyneCult™-derived organoids exhibited a representative mix of secretory and ciliated cells as it occurs in vivo. This demonstrates that GyneCult™ FTOM offers (D) superior growth and (E, F) multilineage representation when compared head-to-head with the DIY formulation. Data was used with permission from the Huntsman Lab, BC Cancer Research Center.

A Subset of High-Grade Serous Ovarian Cancer Organoids Can Be Cultured in GyneCult™ FTOM and Recapitulate Parental Tumor Biology in 3D

Figure 7. A Subset of High-Grade Serous Ovarian Cancer Organoids Can Be Cultured in GyneCult™ FTOM and Recapitulate Tumor Biology in 3D

(A) Representative images of a cryopreserved high-grade serous ovarian cancer (HGSOC) sample cultured in GyneCult™ FTOM, without the addition of the 100x supplement. (B) The resulting HGSOC-derived organoids exhibited a distinct morphology from that of healthy fallopian tube organoids, and (C) expressed HGSOC markers, such as PAX8 (green). (D) Throughout their culture, the HGSOC samples displayed robust growth and an increase in cumulative doublings within five passages. GyneCult™ FTOM supports the growth of a subset of HGSOC samples as organoid cultures to model tumor biology.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

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English
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Technical Manual
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Safety Data Sheet 1
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Safety Data Sheet 2
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Safety Data Sheet 3
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Safety Data Sheet 4
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100-1245
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English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area
Workflow Stages
Workflow Stages for Organoids