Donor-to-donor variability in primary human organoid cultures has not been well characterized. As these cultures contain multiple cell types, there is greater concern that variability could lead to increased noise. In this work we investigated donor-to-donor variability in human gut adult stem cell (ASC) organoids. We examined intestinal developmental pathways during culture differentiation in ileum- and colon-derived cultures established from multiple donors, showing that differentiation patterns were consistent among cultures. This finding indicates that donor-to-donor variability in this system remains at a manageable level. Intestinal metabolic activity was evaluated by targeted analysis of central carbon metabolites and by analyzing hormone production patterns. Both experiments demonstrated similar metabolic functions among donors. Importantly, this activity reflected intestinal biology, indicating that these ASC organoid cultures are appropriate for studying metabolic processes. This work establishes a framework for generating high-confidence data using human primary cultures through thorough characterization of variability.

原代人类器官培养物中供体间的变异性尚未得到很好的表征。由于这些培养物包含多种细胞类型，因此更担心变异性可能导致噪音增加。在这项工作中，我们研究了人类肠道成体干细胞 (ASC) 类器官中供体间的变异性。我们检查了从多个供体建立的回肠和结肠衍生培养物中培养分化过程中的肠道发育途径，表明不同培养物的分化模式是一致的。这一发现表明，该系统中供体间的变异性仍处于可控水平。通过对中心碳代谢物的靶向分析和通过分析激素产生模式来评估肠道代谢活性。两项实验都证明了供体之间的代谢功能相似。重要的是，这一活动反映了肠道生物学，表明这些 ASC 类器官培养物适合研究代谢过程。这项工作建立了一个框架，通过对变异性的彻底表征，使用人类原代文化生成高可信度数据。