CORE--TISSUE CULTURE AND MONOCLONAL ANTIBODIES

对导致与发育障碍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通过体外模型系统进行，如细胞培养，包括转染外源基因的细胞和来自转基因和敲除小鼠的细胞。在理解导致智力迟钝的疾病（如唐氏综合症或粘多糖病）的生化病理机制方面取得的进展，严重依赖于研究分离细胞培养系统中改变的细胞和生化环境的技术的发展。此外，最先进的发育生物学研究经常要求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，以及将外源基因转染到细胞中的构建物。组织培养/单克隆抗体实验室（TCMAC）通过提供核心设施和服务，促进了堪萨斯MRDDRC研究人员的研究，使他们能够开发、维护和表征许多细胞模型系统，以探索正常细胞生长的各个方面，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药物的作用。在过去的拨款期间，TCMAC实验室对设施的许多改进感到非常自豪，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，以满足个人研究人员的需求。为Robert Palazzo所做的抗体工作为他识别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极好的工具。为伊莱亚斯Michaelis开发的一系列抗体已经导致了对慢性乙醇暴露的蛋白质表达和定位的重要发现。在过去的资助期间，该机构开发的抗体目前正在其他大学进行合作研究，一家大公司表示有兴趣销售这些高度特异性和有价值的工具，以便在国际层面上为科学家提供这些工具。随着TCMAC展望未来和加强中心对生物行为过程的关注，核心将在涉及基因转染、胚胎干细胞生长和其他生产转基因或基因敲除小鼠所需技术的研究中发挥越来越大的作用。TCMAC一直是并且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工作，鼓励他们将自己的才能应用到中心的新的、高生产力的科学家身上，并鼓励他们将自己的才能应用到与发育障碍有关的问题上，发挥了极其重要的作用。例如，在Rick Dobrowsky， Doug Ruden和Robert Palazzo加入KU后不久，TCMAC就吸引了他们加入MRDDRC。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招聘新教员，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反应器，熟练的技术支持和最先进的设备，以促进和加强MRDDRC研究人员对导致发育障碍的分子事件的研究。TCMAC将继续通过以下方式促进堪萨斯大学MRDDRC的研究：(1)维护一个集中的设施，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，并为需要持续供应抗体的项目维持杂交瘤细胞系。

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.