A novel 3D liver organoid system for elucidation of hepatic glucose metabolism.
A novel 3D liver organoid system for elucidation of hepatic glucose metabolism.
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一种用于阐明肝脏葡萄糖代谢的新型 3D 肝脏类器官系统
DOI：
10.1002/bit.23349
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发表时间：
2012-02
影响因子：
3.8
通讯作者：
Meng, Qin
中科院分区：
文献类型：
--
作者：
Lu, Yanhua;Zhang, Guoliang;Shen, Chong;Uygun, Korkut;Yarmush, Martin L.;Meng, Qin
肝脏葡萄糖代谢是肥胖和糖尿病等疾病以及抗高血糖药物筛选的关键因素。二维构型的肝细胞培养在肝细胞正常发挥功能的体外模型中受到限制，而进行三维（3D）培养的真正实用的平台不可用。在这项工作中，我们提出了一种实用的肝细胞类器官培养方法，用于阐明标称和应激条件下的葡萄糖代谢。采用这种在中空纤维反应器内培养细胞的新方法，观察到肝细胞自组装成3D球形类器官，保留紧密连接，并显示出增加的肝脏特异性功能。与单层培养和夹心培养相比，肝细胞类器官显示出更高的细胞内糖原含量、葡萄糖消耗和肝细胞生成，并且接近体内值，这也通过关键酶的基因表达证实。此外，肝细胞类器官对胰岛素、胰高血糖素和地塞米松的激素激发表现出更真实的敏感性。最后，暴露于高葡萄糖表现出毒性，包括线粒体膜电位的改变，脂质积累，和活性氧的形成，类似于在体内的反应，这是不能捕获的单层培养。总的来说，肝细胞类器官比肝细胞单层和三明治培养物更好地模拟体内功能，这表明其适用于抗高血糖药物筛选等应用。
Hepatic glucose metabolism is a key player in diseases such as obesity and diabetes as well as in antihyperglycemic drugs screening. Hepatocytes culture in two-dimensional configurations is limited in vitro model for hepatocytes to function properly, while truly practical platforms to perform three-dimensional (3D) culture are unavailable. In this work, we present a practical organoid culture method of hepatocytes for elucidation of glucose metabolism under nominal and stress conditions. Employing this new method of culturing cells within a hollow fiber reactor, hepatocytes were observed to self-assemble into 3D spherical organoids with preservation of tight junctions and display increased liver-specific functions. Compared to both monolayer culture and sandwich culture, the hepatocyte organoids displayed higher intracellular glycogen content, glucose consumption, and gluconeogenesis and approached the in vivo values, as also confirmed by gene expression of key enzymes. Moreover, hepatocyte organoids demonstrated more realistic sensitivity to hormonal challenges with insulin, glucagon, and dexamethasone. Finally, the exposure to high glucose demonstrated toxicities including alteration of mitochondrial membrane potential, lipid accumulation, and reactive oxygen species formation, similar to the in vivo responses, which was not captured by monolayer cultures. Collectively, hepatocyte organoids mimicked the in vivo functions better than hepatocyte monolayer and sandwich cultures, suggesting suitability for applications such as antihyperglycemic drugs screening.
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DOI：
10.1016/s0167-4889(99)00080-4
发表时间：
1999-08-12
影响因子：
5.1
作者：
Davies, GF;Khandelwal, RL;Roesler, WJ
通讯作者：
Roesler, WJ
影响因子：
2.6
作者：
Beale, Elmus G.;Harvey, Brandy J.;Forest, Claude
通讯作者：
Forest, Claude
DOI：
10.1007/10_029
发表时间：
2007-01-01
期刊：
TISSUE ENGINEERING II: BASICS OF TISSUE ENGINEERING AND TISSUE APPLICATIONS
影响因子：
--
作者：
Nahmias, Yaakov;Berthiaume, Francois;Yarmush, Martin L.
通讯作者：
Yarmush, Martin L.
DOI：
10.1016/s0022-2143(98)90134-2
发表时间：
1998-12-01
期刊：
JOURNAL OF LABORATORY AND CLINICAL MEDICINE
影响因子：
--
作者：
Ikezawa, Y;Yamatani, K;Sasaki, H
通讯作者：
Sasaki, H
影响因子：
4.9
作者：
Noshiro, O;Hirayama, R;Shikama, H
通讯作者：
Shikama, H