Enhanced O2 Delivery to C3A Hepatocytes

描述(由申请人提供)：生物人工肝辅助装置(BLAD)利用体外人工装置(通常是中空纤维(HF)生物反应器)内的肝细胞来复制广泛的肝功能。然而，在创造一个可行的BLAD的过程中，仍然需要克服几个挑战，包括为肝细胞提供适当的氧合。维持肝细胞的活性和分化功能高度依赖于氧气供应。提供和维持足够的氧气水平的一个有希望的方法是通过向设备内循环的介质中添加氧气载体，如牛红细胞(BRBC)。我们推测，在BLAD中使用bRBC将允许增加氧气输送，从而促进长期的分化功能和细胞团的保存。我们计划通过追求以下两个特定目标来验证这一假设：1)当bRBC(无论是正常的还是交联的)在循环介质中以不同的bRBC浓度和不同的氧气入口分压存在时，测量并关联肝脏HF生物反应器的全局肝细胞耗氧量(GHOC)与肝细胞的增殖、代谢和合成功能；以及2)在不同的氧合情景下测量并关联肝脏HF生物反应器的GHOC与肝细胞的生物转化和解毒功能。这个项目意义重大，因为它详细研究了在HF生物反应器中培养时，氧气水平对肝细胞功能和活力的影响；据我们所知，这一点还不清楚。这些特定目标中的任何一个或两个的成功将有助于实现这一应用的长期目标，即设计一种能够维持多种肝功能的Blad。从这项研究中获得的知识有望对含有HF生物反应器的肝细胞用作潜在的刀片的设计有所裨益。

DESCRIPTION (provided by applicant): The bioartificial liver assist device (BLAD) utilizes hepatocytes, maintained within an extracorporeal artificial device (commonly a hollow fiber (HF) bioreactor), to reproduce a broad range of liver functions. However, several challenges must still be overcome in the creation of a viable BLAD, including providing appropriate oxygenation to the hepatocytes. Maintaining hepatocyte viability and differentiated function is highly dependent on oxygen provision. 1 promising way to deliver and sustain adequate oxygen levels is through the addition of an oxygen carrier, such as bovine red blood cells (bRBCs), to the media circulating within the device. We hypothesize that the use of bRBCs within a BLAD will allow for increased oxygen delivery, thus promoting prolonged differentiated function and preservation of cell mass. We plan to test this hypothesis by pursuing the following 2 specific aims: 1) Measure and correlate the hepatic HF bioreactor global hepatocyte oxygen consumption (GHOC) with hepatocyte proliferation and metabolic and synthetic functions when bRBCs (either normal or cross-linked) are present in the circulating media at varied bRBC concentrations and at varied inlet partial pressures of oxygen; and 2) Measure and correlate the hepatic HF bioreactor GHOC with hepatocyte biotransformation and detoxification functions under differing oxygenation scenarios. This project is significant in that it is a detailed examination of the impact of oxygen levels on hepatocyte function and viability when cultured within a HF bioreactor; which to the best of our knowledge is not well understood. The success of any or both of these specific aims will aid in achieving the long-term goal of this application, which is to design a BLAD that is capable of sustaining a wide variety of liver functions. The knowledge gained from this study is expected to be of benefit in the design of hepatocyte containing HF bioreactors for use as potential BLADs.