Abstract：
Currently, there are many challenges in the culture of human induced pluripotent stem (iPS) cell-derived intestinal organoids (HIOs) for use in drug discovery, disease research, and regenerative medicine. For example, the main culture method, embedding culture, makes industrial large-scale culture difficult, and Matrigel, which is used for almost all HIO cultures, is not respected for its application in regenerative medicine. To overcome these challenges, we herein propose a new culture method using low concentrations of natural polysaccharides in a suspension culture. In the present study, five natural polysaccharides free from heterologous animal-derived components were used, and HIOs were successfully cultured in suspension with FP001 and FP003, which are microbial exopolysaccharide analogs of gellan gum. The fabricated HIOs were similar to living intestinal tracts with respect to their gene expression, microstructure, and protein expression. The observed activities of the drug metabolizing enzymes and drug transporters in the generated HIOs suggested that they have pharmacokinetic functions. We believe that suspension culture of HIOs using FP001 or FP003 can be widely applied to not only drug discovery research but also disease research and regenerative medicine.
摘要：
目前,在用于药物发现的人类诱导多能干细胞（iPS）细胞衍生的肠道类器官（HIOs）的培养中存在许多挑战，疾病研究,和再生医学。例如,主要的培养方法，嵌入文化，使工业大规模文化变得困难，还有Matrigel,它几乎用于所有的HIO文化，不尊重其在再生医学中的应用。为了克服这些挑战，我们在此提出了一种在悬浮培养物中使用低浓度天然多糖的新培养方法。在本研究中,使用了五种不含异源动物来源成分的天然多糖，和HIOs与FP001和FP003悬浮培养成功，FP001和FP003是结冷胶的微生物外多糖类似物。制造的HIOs在基因表达方面与活体肠道相似，微观结构，和蛋白质表达。在生成的HIOs中观察到的药物代谢酶和药物转运蛋白的活性表明它们具有药代动力学功能。我们认为，使用FP001或FP003悬浮培养HIOs不仅可以广泛应用于药物发现研究，还可以用于疾病研究和再生医学。
