Suboptimal uterine fluid (UF) composition can lead to pregnancy loss and likely contributes to offspring susceptibility to chronic adult-onset disorders. However, our understanding of the biochemical composition and mechanisms underpinning UF formation and regulation remain elusive, particularly in humans. To address this challenge, we developed a high-throughput method for intraorganoid fluid (IOF) isolation from human endometrial epithelial organoids. The IOF is biochemically distinct to the extraorganoid fluid (EOF) and cell culture medium as evidenced by the exclusive presence of 17 metabolites in IOF. Similarly, 69 metabolites were unique to EOF, showing asymmetrical apical and basolateral secretion by the in vitro endometrial epithelium, in a manner resembling that observed in vivo. Contrasting the quantitative metabolomic profiles of IOF and EOF revealed donor-specific biochemical signatures of organoids. Subsequent RNA sequencing of these organoids from which IOF and EOF were derived established the capacity to readily perform organoid multiomics in tandem, and suggests that transcriptomic regulation underpins the observed secretory asymmetry. In summary, these data provided by modeling uterine luminal and basolateral fluid formation in vitro offer scope to better understand UF composition and regulation with potential impacts on female fertility and offspring well-being.

次优的子宫液 (UF) 成分可导致流产，并可能导致后代对慢性成人发病的疾病的易感性。然而，我们对支持超滤形成和调节的生化成分和机制的理解仍然难以捉摸，尤其是在人类中。为了应对这一挑战，我们开发了一种从人类子宫内膜上皮类器官中分离类器官内流体 (IOF) 的高通量方法。IOF 在生物化学上与类外液 (EOF) 和细胞培养基不同，IOF 中仅存在 17 种代谢物就证明了这一点。同样，69 种代谢物是 EOF 独有的，显示出体外子宫内膜上皮的不对称顶端和基底外侧分泌，其方式类似于体内观察到的方式。对比 IOF 和 EOF 的定量代谢组学特征，揭示了类器官的供体特异性生化特征。随后对衍生出 IOF 和 EOF 的这些类器官进行 RNA 测序，建立了轻松串联进行类器官多组学的能力，并表明转录组调控支持观察到的分泌不对称。总之，这些通过模拟体外子宫腔和基底外侧液形成提供的数据为更好地了解 UF 成分和调节以及对女性生育能力和后代健康的潜在影响提供了范围。并表明转录组调控支持观察到的分泌不对称。总之，这些通过模拟体外子宫腔和基底外侧液形成提供的数据为更好地了解 UF 成分和调节以及对女性生育能力和后代健康的潜在影响提供了范围。并表明转录组调控支持观察到的分泌不对称。总之，这些通过模拟体外子宫腔和基底外侧液形成提供的数据为更好地了解 UF 成分和调节以及对女性生育能力和后代健康的潜在影响提供了范围。