Holthaus David, Hedemann Nina, Geweniger Sophia Theresa, Le Huy Duc, Alkatout Ibrahim, Kessler Mirjana, Meyer Thomas F
Unit for 3D-Patient Avatars and Personalized Medicine, Department of Gynecology and Obstetrics, Christian-Albrechts-University Kiel & University Hospital Schleswig-Holstein, Kiel, Germany.
Department of Gynecology and Obstetrics, Christian-Albrechts-University Kiel & University Hospital Schleswig-Holstein, Kiel, Germany.
Bio Protoc. 2025 Aug 20;15(16):e5412. doi: 10.21769/BioProtoc.5412.
The female reproductive tract is comprised of different regions, each with distinctive physiological characteristics. One of them is the fallopian tubes, which are vital for human reproductive health and success. The ability to model their function and physiology is of utmost importance. So far, in vitro models have been based on a few immortalized or cancer cell lines derived from fallopian tube cells that lacked differentiated, specialized cell types and did not allow for the study of cancer initiation due to their implicit biases. Organoids, in contrast, overcome these limitations and provide an advanced, three-dimensional system for the study of healthy fallopian tube physiology and pathology. Fallopian tube organoids are comprised of epithelial progenitors that can be enriched using chemical or hormonal treatment into the different cell types that are found in the in vivo tissue, namely detyrosinated-tubulin-positive ciliated cells or paired-box protein 8 (PAX8)-positive secretory cells. This protocol provides a step-by-step guide for the establishment and maintenance of a long-term culture of organoids from healthy human fallopian tube tissue. The organoid model described here closely mimics the in vivo physiology and anatomy of human fallopian tube epithelium and provides a comprehensive basis for future studies on its underlying molecular characteristics and possible pathology. Key features • Provides a step-by-step guide for the establishment of long-term fallopian tube organoid cultures. • Allows for rapid extension of fallopian tube epithelial progenitor cells with a yield of up to 1 × 10 cells within 3 weeks of isolation. • Fallopian tube organoids closely mimic healthy physiology, being comprised of multiple different cell types, like detyrosinated-tubulin-positive ciliated cells or paired-box protein 8 (PAX8)-positive secretory cells. • Further enrichment of secretory cells by hormonal treatment and ciliated cells by chemical treatment is possible.
女性生殖道由不同区域组成，每个区域都有独特的生理特征。其中之一是输卵管，它对人类生殖健康和生育至关重要。模拟其功能和生理的能力至关重要。到目前为止，体外模型一直基于少数源自输卵管细胞的永生化或癌细胞系，这些细胞系缺乏分化的、专门的细胞类型，并且由于其内在偏差而无法用于癌症起始的研究。相比之下，类器官克服了这些限制，并为研究健康输卵管的生理和病理提供了一个先进的三维系统。输卵管类器官由上皮祖细胞组成，这些祖细胞可以通过化学或激素处理富集为体内组织中发现的不同细胞类型，即去酪氨酸化微管蛋白阳性的纤毛细胞或配对盒蛋白8（PAX8）阳性的分泌细胞。本方案提供了一个逐步指南，用于从健康人输卵管组织建立和维持类器官的长期培养。这里描述的类器官模型紧密模拟了人输卵管上皮的体内生理和解剖结构，并为未来对其潜在分子特征和可能病理的研究提供了全面的基础。关键特性 • 提供建立长期输卵管类器官培养的逐步指南。 • 允许快速扩增输卵管上皮祖细胞，在分离后3周内产量可达1×10个细胞。 • 输卵管类器官紧密模拟健康生理，由多种不同细胞类型组成，如去酪氨酸化微管蛋白阳性的纤毛细胞或配对盒蛋白8（PAX8）阳性的分泌细胞。 • 通过激素处理进一步富集分泌细胞，通过化学处理富集纤毛细胞是可能的。
