Commercialization of Human Omental Mesothelial Cells for Research

描述(申请人提供)：间皮细胞已被发现在肿瘤转移、腹膜透析和炎症反应中起关键作用。这些细胞是特殊的上皮细胞，排列在腹膜、心包、胸膜腔和内脏，为入侵生物和物理损害提供了一种屏障。卵巢肿瘤的附着通过癌细胞与间皮细胞结合并迁移到周围组织和血管系统而发生。腹膜透析依赖于腹膜间皮细胞完整的运输功能，以允许废物从潜在的血管系统转移到腹膜腔内的透析液中。宿主对腹膜感染的反应是由腹膜间皮细胞启动的炎性级联反应和细胞因子的释放所介导的。对间皮细胞生物学的研究仍在继续，目标是开发方法，为疾病提供更有效的治疗。目前，美国还没有这些细胞的商业来源，这需要研究人员自己分离和鉴定这些细胞。这一过程非常耗时，需要接触到人体组织，并在细胞的制备和质量方面引入了差异。Zen-Bio将通过向研究界提供具有良好特性的间皮细胞培养系统来满足这一需求。该项目的第一阶段仅专注于生成优化的原代人类间皮细胞系统。其具体目的是优化人大网膜间皮细胞的分离和增殖，然后仔细鉴定其细胞特性。分离和繁殖优化将通过系统地修改现有方案的各个方面来实现，以提高活细胞产量，同时最大限度地减少种群倍增。将对细胞特性进行详细分析，包括在培养中的寿命、细胞特异性基因表达和细胞角蛋白生产，以验证细胞系统。这些分析将构成未来质量控制程序的基础。该项目的第二阶段包含两个具体目标，这两个目标将在Zen-Bio产生额外的合同服务。目的分析间皮细胞转分化(上皮向间充质转化)过程中细胞因子和基因表达的变化。这一目标的完成直接导致目标二，即在Zen-Bio建立间皮细胞合同服务。这些服务包括分析细胞因子分泌、基因表达、肿瘤细胞-间皮细胞相互作用和细胞毒性。该提案预计将提供多种产品：原生人类间皮细胞系统、支持介质和试剂、合同化验服务以及间皮细胞研究试剂盒。在该项目完成后，将向研究人员提供商业上可用的、完全表征的原代人类间皮细胞系统和支持试剂。这一目前尚不可用的系统的可获得性将为研究抑制卵巢肿瘤附着、延长腹膜透析的效用和治疗腹膜炎的新方法提供更广泛的机会。

DESCRIPTION (provided by applicant): Mesothelial cells have been found to be pivotal in tumor metastasis, peritoneal dialysis, and inflammatory response. These cells are specialized epithelial cells that line the peritoneal, pericardial, and pleural cavities and internal organs presenting a barrier to invading organisms and physical damage. Ovarian tumor attachments occur through cancer cells binding to mesothelial cells and migrating into the surrounding tissue and vasculature. Peritoneal dialysis relies on the intact transport function of mesothelial cells to allow transfer of waste products from the underlying vasculature to the dialysis fluid in the peritoneal cavity. Host response to peritoneal infection is mediated by the inflammatory cascade initiated and cytokines release by peritoneal mesothelial cells. Research into the biology of mesothelial cells continues with the goal of developing methods to provide more effective treatments for disease. Currently, there are no commercial sources of these cells in the U.S. which requires researchers to isolate and characterize the cells on their own. This process is time consuming, requires access to human tissue, and introduces variation in the preparation and quality of the cells. Zen-Bio will address this need by providing a well characterized mesothelial cell culture system to the research community. Phase I of this project focuses solely on generating an optimized primary human mesothelial cell system. The Specific Aim is to optimize the isolation and propagation of human omental mesothelial cells followed by careful characterization of the cellular properties. Isolation and propagation optimization will be achieved by systematically modifying aspects of existing protocols to enhance viable cell yields while minimizing population doublings. A detailed analysis of cell properties, including longevity in culture, cell specific gene expression, and cytokeratin production will be performed to validate the cell system. These analyses will form the basis of future quality control procedures. Phase II of this project contains two Specific Aims that will generate additional contract services at Zen-Bio. Aim I is to analyze the cytokine and gene expression changes during mesothelial cell transdifferentiation (epithelial to mesenchymal transition). Completion of this Aim leads directly to Aim II which is to establish mesothelial cell contract services at Zen-Bio. These services include analysis of cytokine secretion, gene expression, tumor cell-mesothelial cell interaction, and cytotoxicity. Multiple product offerings are expected from this proposal: a primary human mesothelial cell system, support media and reagents, contract assay services, and kits for mesothelial cell research. At the completion of this project, a commercially available, fully characterized primary human mesothelial cell system and support reagents will be offered to researchers. The accessibility of this currently unavailable system will provide wider opportunity to investigate novel methods to inhibit ovarian tumor attachment, prolong the utility of peritoneal dialysis, and treat peritonitis.