Exploratory Science Center, Merck & Co., Inc., Cambridge, MA 02141, USA.
Exploratory Science Center, Merck & Co., Inc., Cambridge, MA 02141, USA.
Stem Cell Reports. 2021 Sep 14;16(9):2364-2378. doi: 10.1016/j.stemcr.2021.07.016. Epub 2021 Aug 26.
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.
供体间的变异性在原代人源类器官培养中尚未得到很好的描述。由于这些培养物包含多种细胞类型，因此人们更加担心变异性会导致噪声增加。在这项工作中，我们研究了人肠道成体干细胞（ASC）类器官培养物中的供体间变异性。我们检测了从小肠和结肠衍生的多个供体建立的培养物中肠道发育途径在培养分化过程中的变化，结果表明分化模式在培养物之间是一致的。这一发现表明，该系统中的供体间变异性仍处于可管理水平。通过对中心碳代谢物的靶向分析和分析激素产生模式来评估肠道代谢活性。这两项实验均表明供体间具有相似的代谢功能。重要的是，这种活性反映了肠道生物学，表明这些 ASC 类器官培养物适合研究代谢过程。这项工作通过对变异性进行彻底的特征描述，为使用人原代培养物生成高可信度数据建立了一个框架。
