Commercialization of a Human Myocyte and Adipocyte Co-Culture System

描述（由申请人提供）：在过去二十年中，有充分证据表明肥胖的增加与代谢和心血管疾病的发病率增加有关。为了应对相关的预期寿命缩短和昂贵的健康问题，美国国立卫生研究院成立了一个肥胖研究工作组。这个SBIR应用程序中提出的产品开发直接解决了工作组2004年8月战略计划中概述的需求。该计划的原则之一是支持旨在打破肥胖及其相关疾病之间联系的研究。腹部脂肪增加与葡萄糖稳态缺陷相关，并与外周组织胰岛素敏感性降低相关。骨骼肌是胰岛素刺激下葡萄糖处理的重要部位，也是肥胖个体胰岛素抵抗的重要部位。为了支持这一领域的研究，我们建议开发一种具有良好特征的、广泛商业化的人类原代骨骼肌细胞和脂肪细胞共培养系统。这种细胞系统将使研究人员能够研究直接参与2型糖尿病和代谢性疾病病因学的两种细胞类型之间复杂的相互作用。在过去的10年里，Zen-Bio一直在为研究人员提供商业化培养的原代人类细胞，并且已经建立了完成该提案所需的专业知识。该公司的重点一直是脂肪细胞，然而，Zen-Bio的科学家在许多类型的人类原代细胞（包括成肌细胞）的分离、扩增和分化方面拥有专业知识。这一建议的第一阶段包括两个目标。Aim I有望建立健全的人类卫星细胞分离、培养和成肌细胞分化方案。Aim II将在Aim I的基础上，通过优化和验证使用人类脂肪细胞和人类骨骼肌细胞的共培养系统。目标1将通过系统地修改现有的方案来满足商业肌细胞系统的需要。每种细胞类型在共培养条件下的详细活性分析将有助于在Aim II中优化系统。通过监测骨骼肌细胞胰岛素信号转导，验证两种细胞类型在共培养中的相互作用。该项目的第二期包括三个目标。目标1是在商业化之前开发必要的骨骼肌细胞质量控制分析，目标2是开发一套基于骨骼肌细胞的合同分析服务，适用于代谢疾病的研究，目标3是建立一个包含来自同一患者的培养脂肪细胞和骨骼肌细胞的细胞库。该提案预计将提供多种产品：人类骨骼肌细胞系统、使用脂肪细胞和骨骼肌细胞的共培养系统、合同检测服务和骨骼肌细胞检测试剂盒。在该项目完成后，将向研究人员提供市售的、完全经过验证的人类骨骼肌细胞系统、骨骼肌细胞/脂肪细胞共培养系统以及相关的检测试剂盒。这些系统的可用性将为研究代谢性疾病的新方法提供机会，并为检查这两种细胞类型之间复杂的相互作用提供独特的方法。

DESCRIPTION (provided by applicant): The well documented rise in obesity over the past two decades correlates with increased incidence rates of metabolic and cardiovascular disease. In response to the associated reduced life expectancy and expensive health problems the NIH has established an Obesity Research Task Force. The product development proposed in this SBIR application directly addresses the needs outlined in the Task Force's August 2004 Strategic Plan. One of the tenets of the plan is supporting research toward breaking the link between obesity and its related disorders. Increased abdominal adiposity correlates with defects in glucose homeostasis and is associated with decreased insulin sensitivity in peripheral tissue. Skeletal muscle is an important site of insulin-stimulated glucose disposal and is often the site of insulin resistance in obese individuals. To support research in this area, we propose to develop a well characterized, widely commercially available co-culture system of primary human skeletal myocytes and adipocytes. This cell system will allow researchers to investigate the complex interaction between two cell types directly involved in the etiology of Type 2 diabetes and metabolic disease. Zen-Bio has been providing cultured primary human cells commercially to researchers for the past 10 years and has established the expertise required for the completion of this proposal. The focus of the company has been adipocytes, however, scientists at Zen-Bio have expertise in the isolation, expansion, and differentiation of many types of human primary cells, including myoblasts. Phase I of this proposal consists of two aims. Aim I is expected to establish robust human satellite cell isolation, culturing, and myoblast differentiation protocols. Aim II will build on Aim I by optimizing and validating a co-culture system using human adipocytes and human skeletal muscle cells. Aim I will be achieved by systematically modifying existing protocols to suit the needs of a commercial myocyte system. Detailed activity analysis of each cell type in co-culture conditions will aid in optimizing the system in Aim II. Validation of the interaction between the two cell types in co-culture will be assessed by monitoring skeletal muscle cell insulin signal transduction. Phase II of the project consists of three aims. Aim I is to develop the necessary quality control assays for skeletal muscle cells prior to commercialization, Aim II is to develop a suite of skeletal muscle cell based contract assay services suitable for the study of metabolic disease, and Aim III is the establishment of a cell repository containing cultured adipocytes and skeletal muscle cells from the same patient. Multiple product offerings are expected from this proposal: a human skeletal muscle cell system, a co-culture system using adipocytes and skeletal muscle cells, contract assay services, and assay kits for skeletal muscle cells. At the completion of this project, a commercially available, fully validated human skeletal myocyte system, skeletal myocyte / adipocyte co-culture system, and related assay kits will be offered to researchers. The availability of these systems will provide opportunities for new approaches in the investigation of metabolic disease and a unique methodology to examine the complex interaction between these two cell types.