A microfluidically perfused three dimensional human liver model
A microfluidically perfused three dimensional human liver model
复制标题
DOI：
10.1016/j.biomaterials.2015.08.043
复制
发表时间：
2015-12-01
期刊：
影响因子：
14
通讯作者：
Mosig, Alexander S.
中科院分区：
文献类型：
--
作者：
Rennert, Knut;Steinborn, Sandra;Mosig, Alexander S.
在肝脏中，非实质细胞（NPC）在肝细胞极化的调节和代谢功能的维持中起着关键作用。我们在这里报告的建立，整合在血管层的内皮细胞和组织巨噬细胞和肝细胞共培养的星状细胞与肝细胞的肝层的NPC的肝类器官。三维肝脏类器官嵌入微流体灌注的生物芯片中，该芯片能够提供足够的营养供应，并且类似于人类肝脏窦状隙的形态学方面。它利用悬浮膜作为细胞基质，模仿Disse空间。基于发光的传感器点被集成到芯片中，以允许在线测量细胞耗氧量。微流体流的应用诱导ZO-1、转铁蛋白、ASGPR-1的限定表达，沿着肝类器官内MRP-2转运蛋白的表达增加。此外，灌注伴随着5（6）-羧基-2 '，7'-二氯荧光素的肝胆分泌增加和肝细胞微绒毛的形成增强。由此我们得出结论，灌注的肝类器官与人类肝脏具有相关的形态和功能特征，并代表了一种新的体外研究工具，可以在接近生理情况的条件下在细胞水平上研究人类肝细胞生理学。(C)2015爱思唯尔有限公司版权所有。
Within the liver, non-parenchymal cells (NPCs) are critically involved in the regulation of hepatocyte polarization and maintenance of metabolic function. We here report the establishment of a liver organoid that integrates NPCs in a vascular layer composed of endothelial cells and tissue macrophages and a hepatic layer comprising stellate cells co-cultured with hepatocytes. The three-dimensional liver organoid is embedded in a microfluidically perfused biochip that enables sufficient nutrition supply and resembles morphological aspects of the human liver sinusoid. It utilizes a suspended membrane as a cell substrate mimicking the space of Disse. Luminescence-based sensor spots were integrated into the chip to allow online measurement of cellular oxygen consumption. Application of microfluidic flow induces defined expression of ZO-1, transferrin, ASGPR-1 along with an increased expression of MRP-2 transporter protein within the liver organoids. Moreover, perfusion was accompanied by an increased hepatobiliary secretion of 5(6)-carboxy-2',7'-dichlorofluorescein and an enhanced formation of hepatocyte microvilli. From this we conclude that the perfused liver organoid shares relevant morphological and functional characteristics with the human liver and represents a new in vitro research tool to study human hepatocellular physiology at the cellular level under conditions close to the physiological situation. (C) 2015 Elsevier Ltd. All rights reserved.