GyneCult™子宫内膜类器官培养基

用于扩增和分化人子宫内膜类器官的无血清、无酚红、无性类固醇激素培养基。

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用于扩增和分化人子宫内膜类器官的无血清、无酚红、无性类固醇激素培养基。

产品优势

生成对激素具有响应性的类器官，用于模拟月经周期、子宫内膜病变和早期着床
保留关键上皮细胞标志物（PAEP、PAX8、SOX9），构建具有生理相关性的体外子宫内膜模型
可从不同月经周期阶段的组织样本、子宫内膜异位病灶以及上皮细胞含量较低的样本中成功建立类器官培养。
采用无血清配方，可在不同供体样本之间实现稳定一致的类器官扩增，并有效降低实验变异性。

产品组分包括

GyneCult™子宫内膜类器官基础培养基，90 mL
GyneCult™子宫内膜类器官培养基10X添加剂，10 mL
GyneCult™子宫内膜类器官培养基100X添加剂，1 mL

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概览

使用GyneCult™子宫内膜类器官培养基（EOM）生成对激素具有响应性的子宫内膜类器官。该培养基不含血清、酚红和性类固醇激素，专为支持女性生殖生物学研究而设计。在培养过程中，可向GyneCult™ EOM中添加不同浓度的雌激素和/或孕激素，以灵活模拟月经周期的不同阶段。生成的类器官对激素有反应，并包含体内主要上皮细胞类型和标志物，例如孕激素相关子宫内膜蛋白（PAEP）、乙酰化微管蛋白、PAX8和SOX9。

该优化的培养基支持从多种样本来源（包括上皮细胞含量较低的样本）稳定地建立类器官，并且与来自增殖期和分泌期子宫内膜、子宫内膜异位症病灶和经期样本的原代细胞兼容。该培养基成分明确、可生成生理相关的细胞类型、样本来源广泛，可在受控的激素调节下灵活地模拟健康和病变的子宫内膜状态，有助于填补目前子宫内膜和女性生殖健康研究方面的不足。

实验数据

Figure 1. GyneCult™ Endometrial Organoid Medium Allows You to Model Endometrial Biology and the Role of Estrogen and Progesterone

(A) The human endometrium (pink) is a hormone-regulated tissue that responds to the rising and ebbing levels of two critical sex steroid hormones, estrogen (E2) and progesterone (P4). At the beginning of the menstrual cycle, around Day 5, E2 levels steadily rise, causing a deepening of endometrial glands and leading to the thickening of the endometrium in the proliferative phase. Ovulation occurs after the peak of E2 levels, around Day 14 of the menstrual cycle. Following ovulation, the corpus luteum, a temporary collection of cells that forms on the ovary, produces increasing amounts of P4 and moderate levels of E2, initiating the secretory phase and maintaining the endometrial lining. As the corpus luteum degenerates near Day 28, E2 and P4 levels decline, triggering endometrial cell apoptosis and menstruation, during which the functional layer of the endometrium is shed. (B) In vivo, endometrial glands arise from the myometrium and are composed of both a glandular and luminal epithelium that contain secretory and ciliated cell types—both of which can be captured by organoids generated by GyneCult™ Endometrial Organoid Medium (EOM). Depending on the phase of the menstrual cycle, ciliated cells range from 1 - 20% of endometrial cells, peaking on Days 12 - 14, at ovulation. Figures adapted from publications by Critchley et al., 2020 and Chumduri and Turco, 2021 licensed under Creative Commons Attribution CC-BY 4.0.

Figure 2. Overview of the GyneCult™ Endometrial Organoid Medium Workflow for the Robust Generation of EOs

(A) Workflow showing the generation of endometrial organoids (EOs). Cells are seeded in Matrigel® domes at Day 0 at 4000 cells per 20 μL Matrigel®, with up to 3 domes per well in a 24-well non-tissue culture-treated plate. The domes are overlaid in a submerged culture with 500 μL of complete GyneCult™ Endometrial Organoid Medium (EOM) supplemented with 0.2 nM estrogen to facilitate passaging. Full-medium changes in each well were performed every 3 - 4 days, until Day 10 when cultures are ready for passage. (B) Representative images of EO morphologies throughout the GyneCult™ EOM workflow. Organoids with visible lumens are observable under the microscope by Day 7. By Days 10 - 12, cilia beating may be observed under high magnification brightfield microscopy. Further characterization of the organoids to identify secretory or ciliated cells via immunocytochemical (ICC) staining can be performed after Day 10. Scale bar = 500 μm.

Figure 3. GyneCult™ Endometrial Organoid Medium Enables Robust Expansion of EOs Across Donor Samples, Including Epithelial-Sparse Starting Material

(A) Brightfield images of endometrial organoid (EO) cultures from 3 individual donors grown in GyneCult™ Endometrial Organoid Medium supplemented with 0.2 nM estrogen. Cultures were imaged at Seeding (Passage 0, Day 0) and at Passages 1 through 5 (P1 - P5), corresponding to 8 - 10 days post-seeding for each passage. Donors 1 and 3 provided epithelial-rich samples that generated robust organoid cultures in P1. Donor 2, however, provided epithelial-sparse material, as indicated by the presence of elongated cells (stromal) and a lack of epithelial organoids seen in P1 prior to subculture. Scale bar = 500 μm. (B) Cumulative doublings were recorded at each passage as a measure of proliferative capacity. Cultures derived from Donor 2 expanded at comparable rates to epithelial-rich samples, and all cultures achieved at least 10 cumulative doublings over five passages. These results demonstrate that GyneCult™ Endometrial Organoid Medium supports robust expansion of EOs, even from low-epithelial-content starting material.

Figure 4. GyneCult™ Endometrial Organoid Medium Enables Robust EO Formation from Diverse Primary Cell Sources

(A, B, D, E) Representative brightfield images of 2-week endometrial organoids (EOs) cultured in GyneCult™ Endometrial Organoid Medium supplemented with estrogen. Organoids were generated from a range of primary endometrial sources, including tissue sampled from (A) proliferative and (B) secretory endometrium, (D) endometriosis lesions, and from (E) menstrual effluent. (C) E-Cadherin (red) immunostaining confirms the epithelial identity of the organoids. These results demonstrate the versatility of GyneCult™ Endometrial Organoid Medium as a platform for modeling endometrial biology and diseases such as endometriosis. Images courtesy of Dr. Thomas Andrieu from the University of Bern. Scale bar = 100 μm.

Figure 5. EOs Cultured in GyneCult™ Endometrial Organoid Medium Maintain Major Cell Types and Hormone Responsiveness

Immunocytochemistry staining and brightfield images of endometrial organoids (EOs) cultured under distinct hormonal conditions corresponding to the four phases of the menstrual cycle over a 14-day period. A subset of organoids was fixed for imaging at each stage, while remaining cultures progressed to the next phase. (A, E, I) Cultures were maintained in early proliferative conditions (0.2 nM estrogen) for the first 4 days after sample seeding, and transitioned to (B, F, J) late proliferative conditions (1 nM estrogen) in the subsequent 3 days. On Day 7, proliferative phase cultures were harvested and fixed. The rest of the EOs were then cultured under (C, G, K) early secretory conditions (0.2 nM estrogen, 4 nM progesterone) for the next 3 days, and then transitioned to (D, H, L) late secretory (0.5 nM estrogen, 40 nM progesterone) conditions for the remaining 4 days of culture. EOs were stained for (A - D) progestagen-associated endometrial protein (PAEP; green) and acetylated tubulin (red), where apical acetylated-tubulin staining facing the lumen indicates ciliated cells. PAEP, a secreted protein, shows accumulation in the proliferative phases, followed by apical localization and a decrease in cytoplasmic levels in the late secretory phase, suggesting active secretion in organoid cultures. (E - H) Transcription factors PAX8 (green) and SOX9 (red) staining shows that GyneCult™ Endometrial Organoid Medium gives rise to organoid cultures that maintain their stem cell niches (PAX8+SOX9+ cells) and respond to hormones to differentiate into secretory cells (PAX8+SOX9-), while maintaining ciliated cells throughout culture.

产品说明书及文档

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