CORE--TISSUE CULTURE AND MONOCLONAL ANTIBODIES

核心——组织培养和单克隆抗体

• 批准号: 6650623
• 负责人: MARY L. MICHAELIS
• 金额: $ 16.03万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6650623
• 关键词: biomedical facility; mental retardation; molecular pathology; monoclonal antibody; tissue /cell culture

Investigations into the cellular and molecular mechanisms that lead to abnormalities associated with developmental disabilities and mental retardation are increasingly being carried out with in vitro model systems such as cell cultures, including cells transfected with foreign genes and cells derived from transgenic and knockout mice. Advances in understanding the mechanisms for the biochemical pathology of diseases that lead to mental retardation, such as Down syndrome or the mucopolysaccharidoses, have been critically dependent upon the development of techniques for studying the altered cellular and biochemical milieu in isolated cell culture systems. In addition, state-of- the-art research in developmental biology frequently requires that investigators develop unique biological reagents such as polyclonal and monoclonal antibodies, various types of nucleotide probes, and constructs for transfection of foreign genes into cells. The Tissue Culture/Monoclonal Antibodies Laboratory (TCMAC) has facilitated the research of several of the Kansas MRDDRC investigators by providing core facilities and services that enable them to develop, maintain, and characterize numerous cellular model systems for exploring aspects of normal cell growth, signaling pathways that regulate cell viability, and effects of exogenous agents that enhance or disrupt cell growth. The TCMAC Laboratory takes great pride in the many enhancements that have been made in the facility during the past grant period as these improvements enable the lab to provide the highest level of support for MRDDRC investigators. The facility has also successfully developed unique polyclonal and monoclonal antibodies that served the needs of individual investigators. The antibody work done for Robert Palazzo has provided an excellent tool in his efforts to identify critical proteins in the centrosome and monitor changes during the cell cycle. The series of antibodies developed for Elias Michaelis have led to important discoveries regarding protein expression and localization in response to chronic ethanol exposure. Antibodies developed by the facility during the past grant period are currently being utilized at other universities in collaborative studies, and a major company has expressed interested in marketing some of those highly specific and valuable tools so they will be available to scientists on an international level. As the TCMAC looks to the future and means of enhancing the Center's focus on biobehavioral processes, the Core will play an expanding role in studies involving gene transfections, growth of ES cells, and other techniques required to produce transgenic or knockout mice. The TCMAC has been and continues to be a relatively small core, but it has played an extremely important role in attracting new, highly productive scientists to the Center itself and encouraging them to apply their talents to new, highly productive scientists to the Center itself and encouraging them to apply their talents to question related to developmental disabilities. For example, the TCMAC was instrumental in the attracting Rick Dobrowsky, Doug Ruden and Robert Palazzo to the MRDDRC shortly after they joined to KU faculty. This core is likely to continue in this role as the University hires new faculty members in the areas of molecular genetics, bioinformatics and developmental neurobiology. The goal of the TCMAC has been and will continue to be the provision of unique bioreagents, skilled technical support, and state-of-the-art equipment to facilitate and enhance the research of MRDDRC investigators who study molecular events leading to developmental disabilities. The TCMAC will continue to facilitate MRDDRC research at The University of Kansas by: (1) Maintaining a centralized facility that provides a cost-effective alternative to purchase of expensive equipment and services by MRDDRC investigators. (2) Producing and characterizing unique monoclonal/polyclonal antibodies and maintaining hybridoma cell lines for projects requiring a consistent supply of antibodies.

对导致与发育残疾和智力迟钝相关的异常的细胞和分子机制的研究越来越多地使用体外模型系统进行，例如细胞培养物，包括用外源基因转染的细胞和来自转基因和敲除小鼠的细胞。在理解导致精神发育迟滞的疾病（如唐氏综合征或粘多糖沉积症）的生化病理学机制方面的进展，一直严重依赖于在分离细胞培养系统中研究改变的细胞和生化环境的技术的发展。此外，发育生物学的最新研究经常需要研究人员开发独特的生物试剂，如多克隆和单克隆抗体，各种类型的核苷酸探针，以及用于将外源基因转染到细胞中的构建体。组织培养/单克隆抗体实验室（TCMAC）为堪萨斯MRDDRC研究人员的研究提供了便利，提供了核心设施和服务，使他们能够开发、维护和表征多种细胞模型系统，以探索正常细胞生长、调节细胞活力的信号通路以及增强或破坏细胞生长的外源性因子的影响。TCMAC实验室非常自豪地在过去的资助期间在该设施中进行了许多改进，因为这些改进使实验室能够为MRDDRC研究人员提供最高水平的支持。该设施还成功开发了独特的多克隆和单克隆抗体，满足了个别研究人员的需求。为Robert Palazzo所做的抗体工作为他鉴定中心体中的关键蛋白质和监测细胞周期中的变化提供了一个很好的工具。为Elias Michaelis开发的一系列抗体导致了关于蛋白质表达和定位的重要发现，以响应慢性乙醇暴露。该设施在过去的资助期间开发的抗体目前正在其他大学的合作研究中使用，一家大公司表示有兴趣销售一些高度特异性和有价值的工具，以便在国际层面上向科学家提供。随着TCMAC展望未来，以及加强中心对生物行为过程的关注，核心将在涉及基因转染、ES细胞生长和生产转基因或基因敲除小鼠所需的其他技术的研究中发挥越来越大的作用。TCMAC一直是并将继续是一个相对较小的核心，但它在吸引新的，高生产力的科学家到中心本身，并鼓励他们将自己的才能应用于新的，高生产力的科学家到中心本身，并鼓励他们将自己的才能应用于与发育障碍有关的问题方面发挥了极其重要的作用。例如，TCMAC是在吸引里克Dobrowsky，道格鲁登和罗伯特Palazzo到MRDDRC后不久，他们加入KU教师工具。随着大学在分子遗传学、生物信息学和发育神经生物学领域聘用新的教员，这一核心可能会继续发挥这一作用。TCMAC的目标一直是并将继续是提供独特的生物制剂，熟练的技术支持和最先进的设备，以促进和加强研究导致发育障碍的分子事件的MRDDRC研究人员的研究。TCMAC将继续通过以下方式促进堪萨斯大学的MRDDRC研究：（1）维持一个集中的设施，为MRDDRC研究人员购买昂贵的设备和服务提供一个具有成本效益的替代方案。(2)生产和鉴定独特的单克隆/多克隆抗体，并为需要稳定供应抗体的项目维护杂交瘤细胞系。