Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 B51, Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa 226-8503, Japan.
Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama 700-8525, Japan.
J Biosci Bioeng. 2014 Jan;117(1):99-106. doi: 10.1016/j.jbiosc.2013.06.015. Epub 2013 Jul 27.
When constructing an in vitro model of liver tissue to mimic the in vivo liver microenvironment, the major challenge is to preserve and maintain the hepatocyte phenotype. The aim of this study was to develop a novel intelligent hybrid sponge for use in a dense co-culture system designed to simulate the liver microenvironment. We prepared a galactosylated chitosan (GCs)/hyaluronic acid (HA) hybrid sponge using a freeze-drying technique for the co-culture of primary hepatocytes and endothelial cells. Subsequently, we investigated the biocompatibility of the GCs/HA scaffold with primary hepatocytes and endothelial cells in terms of cell attachment, morphology, bioactivity, and maintenance of specific liver functions. The GCs/HA-hybrid sponge demonstrated good biocompatibility not only with primary hepatocytes, but also with endothelial cells. In our model, primary hepatocytes exhibited superior bioactivity and higher levels of liver-specific functions in terms of hepatocyte-specific gene expression, urea production, and testosterone metabolism as compared to a monoculture system. We succeeded in constructing a liver tissue-like model using the GCs/HA-hybrid sponge. Therefore, we anticipate that GCs/HA-hybrid sponges may be a promising matrix for the co-culture of hepatocytes and endothelial cells in liver tissue engineering, and might be employed as a novel co-culture model for applications in toxicology and drug metabolism.
当构建模拟体内肝脏微环境的肝组织体外模型时，主要挑战是保存和维持肝细胞表型。本研究旨在开发一种新型智能复合海绵，用于设计模拟肝脏微环境的密集共培养系统。我们使用冷冻干燥技术制备了一种半乳糖化壳聚糖（GCs）/透明质酸（HA）复合海绵，用于原代肝细胞和内皮细胞的共培养。随后，我们从细胞黏附、形态、生物活性和维持特定肝脏功能等方面，研究了 GCs/HA 支架与原代肝细胞和内皮细胞的生物相容性。GCs/HA-杂化海绵不仅与原代肝细胞，而且与内皮细胞都具有良好的生物相容性。在我们的模型中，与单核培养系统相比，原代肝细胞在肝细胞特异性基因表达、尿素生成和睾酮代谢等方面表现出更高的生物活性和更高水平的特定肝脏功能。我们成功地使用 GCs/HA-杂化海绵构建了类似于肝组织的模型。因此，我们预计 GCs/HA-杂化海绵可能是肝组织工程中肝细胞和内皮细胞共培养的有前途的基质，并可能作为毒理学和药物代谢应用的新型共培养模型。
