NOVEL OXYGENATION SYSTEM FOR 3D GROWTH OF LIVER CELLS

这个项目的长期目标是开发一种供氧系统 它可以为肝细胞提供维持长时间所需的氧气 体外长期增殖。这些细胞在体内有足够的氧合 空气中氧气通过空气缓慢扩散，阻碍了培养。 覆盖液体介质。正在开发的系统克服了这一点 限制。细胞生长在全氟碳偶联明胶上 在一层全氟十氢林(PFD)上覆盖底物。这提供了氧气 直接发送到接口处的单元。该底物上的HeLa细胞 生长形成19层以上的组织样结构。 正在进行这些领域的工作，以使这种方法适用于 肝细胞生长：1)复氧PFD系统的建立，2) 适合胶原蛋白的全氟偶联物的开发，以及3) 一种含有所需因子的浓缩培养基的研制 维持核扩散。 这种新的氧合系统将为肝细胞培养提供巨大的 改进了测试药物和研究酒精对人体健康影响的系统 肝脏。肝细胞与其他肝细胞的共培养将提供 异型组织样结构，可用作体外组织 为肝功能受损的患者植入植入物。 建议的商业应用：有三种商业产品 在这个项目中开发的。第一种涉及一种肝细胞培养液 它的表现超过了商业上可用的媒体。第二个问题涉及新的 共轭全氟碳化合物，将用于蛋白质结合。 最后也是最重要的一部分包括前两个产品以及一个 细胞培养室，将被用来以3D方式生长肝细胞。这些 细胞可用于研究正常人的酒精和药物代谢 肝细胞。

The long term goal of this project is to develop an oxygenation system which can provide hepatocytes with the oxygen they need to maintain long term proliferation in vitro. Adequate oxygenation of these cells in culture is prevented by the slow rate oxygen diffuses from air through the overlaying liquid medium. The system being developed overcomes this limitation. Cells are grown on a perfluorocarbon conjugated gelatin substratum over a layer of perfluorodecalin (PFD). This provides oxygen directly to the cells at the interface. Hela cells on this substratum grew to form a tissue like structure of more than 19 layers. These areas of work are being undertaken to adapt this method for hepatocyte growth: 1) development of a system for re-oxygenation PFD, 2) development of suitable perfluoro-conjugates for collagens, and 3) development of an enriched culture media containing factors needed to sustain proliferation. This new oxygenation system for hepatocyte culture will provide greatly improved systems for testing drugs and studying the effects of alcohol on the liver. Co-culture of hepatocytes with other liver cells will provide heterotypic tissue like structures which could be used as extra-corporeal implants in patients with compromised-liver function. PROPOSED COMMERCIAL APPLICATION: There are three commercial products being developed in this project. The first involves a liver cell culture media which outperforms commercially available media. The second involves new conjugated perfluorocarbons which will be used for protein attachment. The last and most important includes the first two products along with a cell culture chamber which will be used to grow liver cells in 3D. These cells can be used to study alcohol and drug metabolism in normal hepatocytes.