Enhanced O2 Delivery to C3A Hepatocytes

描述（由申请人提供）：生物人工肝辅助装置（BLAD）利用维持在体外人工装置（通常为中空纤维（HF）生物反应器）中的肝细胞，以重现广泛的肝功能。然而，在创建可行的BLAD中仍必须克服几个挑战，包括为肝细胞提供适当的氧合。维持肝细胞活力和分化功能高度依赖于氧供应。输送和维持足够氧气水平的一种有希望的方法是通过向设备内循环的培养基中添加氧载体，如牛红细胞（bRBC）。我们假设在BLAD内使用bRBC将允许增加氧递送，从而促进延长的分化功能和细胞团的保存。我们计划通过追求以下2个具体目标来测试这一假设：1）当bRBC被激活时，测量肝HF生物反应器整体肝细胞耗氧量（GHOC）并将其与肝细胞增殖以及代谢和合成功能相关联。（正常或交联）以不同的bRBC浓度和不同的入口氧分压存在于循环介质中;和2）测量肝HF生物反应器GHOC并将其与不同氧合情况下的肝细胞生物转化和解毒功能相关联。该项目的重要性在于，它详细检查了在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，这一点还没有得到很好的理解。这些特定目标中的任何一个或两个的成功将有助于实现本申请的长期目标，即设计能够维持各种肝功能的BLAD。从本研究中获得的知识预计将有助于设计用作潜在BLAD的含肝细胞HF生物反应器。

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.