Study Transportation of Drugs within Newly Established Murine Colon Organoid Systems.
Study Transportation of Drugs within Newly Established Murine Colon Organoid Systems.
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研究新建立的小鼠结肠类器官系统中药物的运输。
DOI：
10.1002/adbi.202300103
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发表时间：
2023
期刊：
影响因子：
3.7
通讯作者：
Wang,Qun
中科院分区：
文献类型：
--
作者：
Davoudi,Zahra;Atherly,Todd;Borcherding,DanaC;Jergens,AlbertE;Wannemuehler,Michael;Barrett,TerrenceA;Wang,Qun
小肠 3D 类器官的开发是药物开发和生物学研究的巨大突破。然而，由于缺乏简单、经济有效的类结肠培养方案，结肠类器官（即类结肠）的开发特别具有挑战性。在这里，肠匀浆被描述为用于维持和复制结肠干细胞的培养基的补充物。通过该培养方案产生的克隆体表现出显着的增殖和分化（3个月）。培养的类结肠和原代结肠组织在结构和功能方面具有相似性。为了评估类集落的功能，使用 4 和 40 kDa 异硫氰酸荧光素葡聚糖 (FITC-DEX) 悬浮液进行渗透性测试。据观察，两者都不能渗透健康的上皮屏障。 P-糖蛋白受体是一种重要的药物外排泵，可减轻潜在的药物毒性，其功能受到操纵，维拉帕米的抑制功能和监测罗丹明 123 的摄取就证明了这一点。此外，影响氯离子转运的毛喉素治疗会导致类器官肿胀；这证实了 CFTR 转运蛋白在结肠中的功能表达。这种生成类集落的方案有望用于高通量药物筛选、毒性测试和口服药物开发。
The development of 3D organoids of the small intestine is a tremendous breakthrough in drug development and biological research. However, the development of colonic organoids (i.e., colonoids) is particularly challenging due to a lack of simple, cost‐effective protocols for colonoid cultivation. Here, intestinal homogenates are described as a supplement to the culture medium for maintaining and replicating colonic stem cells. Colonoids generated by this cultivation protocol demonstrate substantial proliferation and differentiation (3 months). There is a similarity between cultured colonoids and primary colon tissue regarding structure and functionality. To evaluate the functionality of colonoids, permeability testing is performed with suspensions of 4 and 40 kDa fluorescein isothiocyanate‐dextran (FITC‐DEX). It is observed that neither can permeate the healthy epithelial barrier. The P‐glycoprotein receptor, a vital drug efflux pump mitigating potential drug toxicity, is functionally manipulated, as evidenced by its inhibition function by verapamil and monitoring uptake of Rhodamin 123. In addition, Forskolin treatment which affects chloride transport results in organoid swelling; this confirms the functional expression of the CFTR transporter in the colonoids. This protocol to generate colonoids is promising for high‐throughput drug screening, toxicity testing, and oral drug development.