Commercialization of Human Mammary Basal / Luminal Cell System for Research

描述（由申请人提供）：乳腺癌是女性中第二常见的癌症。2007年，美国大约有18万名妇女将被诊断出患有恶性乳腺癌，大约4万人将死亡。大多数乳腺癌起源于产乳腺的小叶或导管细胞。在这些结构中，有两种主要的细胞类型：由基底肌上皮包围的内腔细胞。这些细胞类型是不同的，是各种形式乳腺癌的前体。因此，重要的是独立地以及在共培养模型系统中研究这些细胞群体。目前用于培养人乳腺上皮细胞的方法主要选择基础表型的那些。因此，在美国没有质量受控的、匹配的基底细胞和管腔细胞（来自个体供体）的商业来源，迫使研究人员自己分离和表征细胞。这个过程是耗时的，需要接近人体组织，并在细胞的制备和表征中引入变化。原代培养的人乳腺源性细胞是目前用于研究人类乳腺癌的发生和理解的理想模型。由于原代培养的人乳房来源的管腔和基底细胞的一致商业来源不可用，Zen-Bio，Inc.将通过为研究团体提供一个具有良好特征的系统来满足这一需求。该项目的I期重点是生成优化的原代人乳腺源性基底/管腔细胞系统。具体目的是优化人乳腺上皮细胞的分离和增殖，然后表征细胞表型。将通过系统地修改现有方案的各个方面来实现分离和繁殖的优化，以提高活细胞产量，同时最大限度地减少群体doubility。将对细胞特性进行详细分析，包括培养寿命、细胞特异性基因表达和细胞角蛋白产生，以初步验证细胞系统。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特定目标，将在Zen-Bio产生额外的合同服务。目标I是分析两种细胞类型之间的细胞因子和基因表达差异。完成这一目标直接导致目标II，即在Zen-Bio为学术和制药领域建立合同研究服务。这些服务包括细胞因子分泌、基因表达、肿瘤细胞-脂肪相互作用和细胞毒性的分析。该提案预计将提供多种产品：匹配的原代人乳腺细胞系统、支持培养基和试剂、合同检测服务以及用于基底/管腔细胞研究的试剂盒。公共卫生相关性：在该项目完成时，将向研究人员提供市售的、充分表征的人原代乳腺源性基底/管腔细胞系统和支持试剂。这个目前不可用的系统的可访问性不仅将提供更大的机会来研究新方法以进一步了解乳腺癌的病因，而且还将研究潜在的治疗方法。

DESCRIPTION (provided by applicant): Breast cancer is the second most common cancer among women. Roughly 180,000 women in the U.S. will be diagnosed with malignant breast cancer in 2007-some 40,000 will die. The majority of breast cancers originate in the lobular or ductal cells of the milk-producing glands. In these structures, there are two main cell types: the inner luminal cells surrounded by basal myoepithelium. These cells types are distinct and are the precursors to various forms of breast cancer. As such, it is it is important to study these cell populations independently as well as in co-culture models systems. Current methods for culturing human mammary epithelial cells predominantly select for those of a basal phenotype. Thus, there are no commercial sources of quality-controlled, matched basal and luminal cells (from individual donors) in the U.S., forcing 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 characterization of the cells. Primary cultured human mammary-derived cells are an ideal model currently used to investigate the genesis and understanding of human breast cancer. Because a consistent commercial source of primary human cultured breast-derived luminal and basal cells is unavailable, Zen-Bio, Inc. will address this need by providing a well characterized system to the research community. Phase I of this project focuses on generating an optimized primary human mammary-derived basal/luminal cell system. The Specific Aim is to optimize the isolation and propagation of human mammary epithelial cells followed by characterization of the cellular phenotypes. Optimization of the isolation and propagation 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 initially 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 differences between the two cell types Completion of this Aim leads directly to Aim II which is to establish contract research services at Zen-Bio for those in both the academic as well as pharmaceutical arenas . These services include analysis of cytokine secretion, gene expression, tumor cell(s)-adipose interaction, and cytotoxicity. Multiple product offerings are expected from this proposal: a matched, primary human mammary cell system, support media and reagents, contract assay services, and kits for basal/luminal cell research. PUBLIC HEALTH RELEVANCE: At the completion of this project, a commercially available, fully characterized human, primary mammary- derived basal/luminal cell system and support reagents will be offered to researchers. The accessibility of this currently unavailable system will not only provide a greater opportunity to investigate novel methods to further understand the etiology of breast cancer but also to investigate potential cures.