Methods in Human ES Cell Research

描述（由申请人提供）：这是一个竞争性的续期申请，申请资金用于继续短期课程“人类胚胎干细胞研究方法”，以前称为“干细胞生物学当前协议”，自2002年以来每年在杰克逊实验室提供。本课程将继续于每年8月初在杰克逊实验室的应用基因组学培训中心举行。本课程将继续汇集一些领先的专家在胚胎干细胞技术，培养学生成功的培养，维护和操作的胚胎干细胞。具体而言，本课程将包括：1)培养基的准备，包括必要的质量控制；2)给料层的制备；3)胚胎干细胞传代技术；4)胚胎干细胞冷冻保存；5)胚胎干细胞转染技术；6)胚胎干细胞及其分化衍生物的FACs分析；7)胚状体形成；8)胚胎干细胞的神经分化；9)胚胎干细胞的造血分化；10)胚胎干细胞的心脏分化。该课程将只使用在NIH注册的人类胚胎干细胞系，特别是UC06， WA09和ES03。本课程的一个主要长期目标是分享和改进标准协议，并最终出版一本技术手册，类似于Hogan等人编辑的流行的“操纵小鼠胚胎”手册。事实上，课程大纲目前包含了一个协议的纲要，试剂清单和研究论文，为干细胞研究人员提供了一个有价值的参考资源。这些材料可根据杰克逊实验室的要求提供。这些目标将通过向16名因其杰出的研究潜力而被选中的参与者提供为期5天的强化课程来实现。2006年，该课程将于8月6日至11日在缅因州巴尔港的杰克逊实验室举行，紧接着是第二届国际干细胞倡议研讨会，届时来自世界各地主要胚胎干细胞研究中心的人类胚胎干细胞研究人员将齐聚一堂，讨论使用共享试剂和方法进行的实验结果。课程结束后，将紧接着在Mount Desert Island生物实验室举行的“陆地和海上的干细胞”研讨会，该研讨会将探讨比较干细胞生物学中更广泛的问题。本课程鼓励学生免费参加这些配套活动。与公共卫生的相关性：目前人类胚胎干细胞成为生物医学研究界普遍存在的研究工具的主要瓶颈仍然是缺乏必要的实践培训来学习如何培养这些敏感细胞。本应用程序中提出的课程将继续为研究人员提供如何在体外培养，操作和分化人类胚胎干细胞的实践培训。

DESCRIPTION (provided by applicant): This is a competing renewal requesting funds for the continuance of the short course "Methods in Human ES Cell Research" formerly known as "Current Protocols in Stem Cell Biology" which has been offered annually at The Jackson Laboratory since 2002. This course will continue to be held in The Jackson Laboratory's Applied Genomics Training Center in early August of each year. This course will continue to bring together some of the leading experts on ES cell technology to train students in the successful culture, maintenance and manipulation of ES cells. Specifically, this course will cover: 1) The preparation of media, including necessary quality control; 2) The preparation of feeder layers; 3) ES cell passaging techniques; 4) ES cell cryopreservation; 5) ES cell transfection techniques; 6) FACs analysis of ES cells and their differentiated derivatives; 7) Embryoid body formation; 8) Neural Differentiation of ES cells; 9) Hematopoietic Differentiation of ES cells; and 10) Cardiac Differentiation of ES cells. The Course will utilize only human ES cell lines registered with the NIH, specifically UC06, WA09 and ES03. One major long term goal for this course will be to share and improve standard protocols, and eventually to publish a techniques manual, similar in style to the popular "Manipulating the Mouse Embryo" manual edited by Hogan et al. In fact, the course syllabus currently contains a compendium of protocols, reagent lists and research papers, providing stem cell researchers with a valuable reference resource. These materials are available upon request from The Jackson Laboratory. These aims will be accomplished by offering an intensive 5-day course to 16 participants chosen for their outstanding research potential. In 2006, the course will be held on August 6-11 at The Jackson Laboratory in Bar Harbor, Maine, and will be immediately preceded by the 2nd Annual International Stem Cell Initiative Workshop at which human ES cell researchers from the major ES cell research centers around the world will meet to discuss the outcome of experiments carried out using shared reagents and methods. The course will be immediately followed by a companion symposium "Stem Cells on Land and at Sea", also held annually, at The Mount Desert Island Biological Laboratory which examines broader issues in comparative stem cell biology. Students in the course are encouraged to attend these companion events free of charge. Relevance to Public Health: The current major bottleneck to human ES cells becoming a ubiquitous research tool in the biomedical research community continues to be the scarcity of the hands-on training necessary to learn how to culture these sensitive cells. The course proposed in this application will continue to provide hands-on training for investigators in how to culture, manipulate, and differentiate human ES cells in vitro.