Human hepatocyte functions in a crossed hollow fiber membrane bioreactor
Human hepatocyte functions in a crossed hollow fiber membrane bioreactor
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DOI：
10.1016/j.biomaterials.2009.01.011
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发表时间：
2009-05-01
期刊：
影响因子：
14
通讯作者：
Drioli, Enrico
中科院分区：
文献类型：
--
作者：
De Bartolo, Loredana;Salerno, Simona;Drioli, Enrico
肝组织工程的一个重要挑战是开发能够促进肝脏重建和调节肝细胞行为的生物人工系统。为了支持人肝细胞的长期维持和分化，研制了一种交叉中空纤维膜生物反应器。该生物反应器由改性聚醚醚酮（PEEK-WC）和聚醚砜（PES）两种不同分子量截止（MWCO）和理化性质的中空纤维（HF）膜交替交叉组装而成，分别作为流入介质和流出介质。这两种纤维组的结合产生了细胞粘附的毛细血管外网络，并通过培养基的交叉流动产生了高质量的交换。对肝脏特异性产物（如白蛋白和尿素）与药物（如地西泮）的运输进行了建模，并与实验代谢数据进行了比较。与实验数据相比，白蛋白的理论代谢物浓度相差7.5%，尿素的理论代谢物浓度相差5%。优化的生物反应器灌注条件使肝脏在尿素合成、白蛋白分泌和地西泮生物转化方面的功能得以维持至培养18天。尿素合成速率高（28.7 μ g/h 10(6)个细胞），地西泮生物转化速率高，证实了该生物反应器的良好性能。在生物反应器中，人肝细胞高水平表达参与地西泮代谢的单个细胞色素P450同工酶。结果表明，交叉HF膜生物反应器能够支持原代人肝细胞的维持，并在所有研究期间保持其肝脏特异性功能。该装置可作为替代动物实验的肝脏组织工程中药物代谢/毒性试验和疾病发病机制研究的潜在工具。2009爱思唯尔有限公司版权所有，
An important challenge in liver tissue engineering is the development of bioartificial systems that are able to favour the liver reconstruction and to modulate liver cell behaviour.A crossed hollow fiber membrane bioreactor was developed to support the long-term maintenance and differentiation of human hepatocytes. The bioreactor consists of two types of hollow fiber (HF) membranes with different molecular weight cut-off (MWCO) and physico-chemical properties cross-assembled in alternating manner: modified polyetheretherketone (PEEK-WC) and polyethersulfone (PES), used for the medium inflow and outflow, respectively. The combination of these two fiber set produces an extracapillary network for the adhesion of cells and a high mass exchange through the cross-flow of culture medium. The transport of liver specific products such as albumin and urea together with the transport of drug such as diazepam was modelled and compared with the experimental metabolic data. The theoretical metabolite concentration differed 7.5% for albumin and 5% for urea with respect to experimental data. The optimised perfusion conditions of the bioreactor allowed the maintenance of liver functions in terms of urea synthesis, albumin secretion and diazepam biotransformation tip to 18 days of culture. In particular the good performance of the bioreactor was confirmed by the high rate of urea synthesis (28.7 mu g/h 10(6) cells) and diazepam biotransformation. In the bioreactor human hepatocytes expressed at high levels the individual cytochrome P450 isoenzymes involved in the diazepam metabolism. The results demonstrated that crossed HF membrane bioreactor is able to support the maintenance of primary human hepatocytes preserving their liver specific functions for all investigated period. This device may be a potential tool in the liver tissue engineering for drug metabolism/toxicity testing and study of disease pathogenesis alternatively to animal experimentation. (C) 2009 Elsevier Ltd. All rights reserved,