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18万女性被诊断为恶性乳腺癌，其中4万人将死亡。大多数乳腺癌起源于产乳腺的小叶细胞或导管细胞。在这些结构中，有两种主要的细胞类型：被基底肌上皮包围的内腔细胞。这些细胞类型不同，是各种乳腺癌的前体。因此，独立研究这些细胞群以及在共培养模型系统中是很重要的。目前培养人乳腺上皮细胞的方法主要选择基础表型的细胞。因此，在美国没有质量控制的、匹配的基底细胞和腔细胞（来自个人供体）的商业来源，迫使研究人员自己分离和表征细胞。这个过程耗时，需要接触人体组织，并且在细胞的制备和表征中引入了变化。原代培养的人乳腺源性细胞是目前用于研究人类乳腺癌发生和了解的理想模型。由于没有一致的人类培养乳腺源性管腔细胞和基底细胞的商业来源，Zen-Bio公司将通过向研究界提供一个特性良好的系统来满足这一需求。该项目的第一阶段重点是产生一个优化的原发性人乳腺源性基底/腔细胞系统。具体目的是优化人乳腺上皮细胞的分离和繁殖，随后表征细胞表型。优化分离和繁殖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加倍。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。这些分析将构成今后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，将在Zen-Bio产生额外的合同服务。目标一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二，即在禅生物为学术和制药领域的人建立合同研究服务。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、支持介质和试剂、合同分析服务以及用于基础/腔内细胞研究的试剂盒。公共卫生相关性：在该项目完成后，将向研究人员提供一种市售的、完全表征的人原代乳腺来源的基底/腔细胞系统和支持试剂。这种目前无法使用的系统的可及性不仅将提供更大的机会来研究进一步了解乳腺癌病因的新方法，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.