CellGuard - A Novel Approach for Protecting Cells During Bioprocessing

描述(申请人提供)：CPSI(细胞保存服务公司)是一家生物技术公司，致力于开发细胞-分子生物学和医学领域的新技术。CPSI的核心分子策略专注于理解和操纵细胞生存和死亡途径，这些途径似乎是细胞特有的，并因细胞操纵而被激活。CPSI对这些应激途径的了解导致了对一系列未来产品的开发，这些产品旨在改善细胞的生物处理。该研究项目的目标是开发第一代CellGuardTM，这是一种组织培养添加剂，旨在减轻细胞在生物处理过程中暴露在不断变化的环境中的反应。CellGuardTM的目标是提供更稳定的培养环境，从而延长细胞在体外的可用间隔，同时将细胞性能保持在标准的、可预测的反应窗口内。为此，CPSI已经确定了一种独特的化合物，FR-48，这是一种显示出希望的化合物。因此，这些化合物为支持CPSI的细胞处理程序提供了巨大的潜力。这项建议的目的是(1)更好地了解FR-48靶向的细胞应激途径，(2)评估FR-48类似物以评估其有效性，以及(3)调查FR-48类似物在操作前、操作中和操作后暴露设置中的使用情况。样品将在三种不同的环境应激状态下进行评估，包括缺氧、营养剥夺和细胞生长/老化。第一阶段的研究将利用HepG2/C3A肝细胞系和原代肝细胞。第二阶段的研究将扩展到更多的细胞类型和更深入的模拟和细胞功能分析。第一阶段的研究将包括使用荧光分析、流式细胞仪、微阵列分析、蛋白质印迹等方法对FR-48‘S作用进行分子分析。该项目的成功完成将导致改进人类肝细胞和其他细胞系统的活性批量细胞培养(生物处理)方案。第二阶段的研究将集中于CellGuardTM产品线的进一步开发和应用，以改善一系列细胞系统的生物处理应用。公共卫生相关性：该项目旨在开发一系列改进的解决方案和协议，以在生物处理过程中保护人类细胞。这种新的方法，统称为CellGuardTM，将导致人类细胞(如肝细胞)的活性和功能的改善。这反过来将提高未来以细胞为基础的药物的质量，并对人类细胞在细胞治疗中的使用产生积极影响。

DESCRIPTION (provided by applicant): CPSI (Cell Preservation Services, Inc) is a biotechnology company dedicated to developing novel technologies in the area of cell-molecular biology and medicine. CPSI's core molecular strategy is focused on understanding and manipulating the cell survival and death pathways that appear to be cell-specific and are activated as a consequence of cell manipulation. CPSI's knowledge of these stress pathways has led to insight into the development of a series of future products designed to improve cell bioprocessing. The goal of this research project is to develop the first generation of CellGuardTM, tissue culture additives designed to mitigate cell response to the continually changing environment a cell is exposed to during a bioprocessing procedure. The objective of CellGuardTM is to provide for a more stable culture environment thereby extending the usable interval of cells in vitro while maintaining cell performance within a standard, predictable response window. To this end, CPSI has identified a unique class of compounds, FR-48, that has shown promise. As such, these compounds offer great potential to support CPSI's cell processing program. The intent of this proposal is to (1) better understand the cell stress pathway targeted by FR-48, (2) evaluate FR-48 analogues to assess their efficacy and (3) investigate the utilization of FR-48 analogs in a pre-, intra, and post-manipulation exposure setting. Samples will be evaluated under three distinct environmental stress regimes including hypoxia, nutrient deprivation, and cell growth/aging. Phase 1 studies will utilize the HepG2/C3A liver cell line and primary hepatocytes. Phase 2 studies will be expanded to additional cell types and more in- depth analogue and cell function analysis. Investigations under this Phase 1 will include molecular analysis of FR-48's action using fluorescent assays, flow cytometry, microarray analysis, western blotting, etc. Successful completion of the project will lead to improved protocols for the active, bulk cell culture (bioprocessing) of human hepatocytes as well as other cell systems. Phase 2 studies will focus on the further development and application of the CellGuardTM product line to improve bioprocessing applications for an array of cell systems. PUBLIC HEALTH RELEVANCE: This project is designed to develop an improved series of solutions and protocols for protecting human cells during bioprocessing. This novel approach, collectively called CellGuardTM, will result in the improved viability and function of human cells such as liver cells. This, in turn, will improve the quality of future cell-based, pharmaceutical drugs as well as positively affect the use of human cells in cell therapy.