Methods in Human ES Cell Research

描述(由申请人提供)：这是一项竞争性的续期申请，要求为杰克逊实验室自2002年以来每年提供的短期课程“人类胚胎干细胞研究的方法”(以前称为“干细胞生物学中的现行方案”)的继续提供资金。该课程将继续于每年8月初在杰克逊实验室的应用基因组学培训中心举行。本课程将继续汇聚一些顶尖的胚胎干细胞技术专家，培训学生如何成功培养、维护和操纵胚胎干细胞。具体地说，本课程将涵盖：1)培养液的制备，包括必要的质量控制；2)饲养层的准备；3)ES细胞的传代技术；4)ES细胞的冷冻保存；5)ES细胞的转染技术；6)ES细胞及其分化的FACS分析；7)拟胚体的形成；8)ES细胞的神经分化；9)ES细胞的造血分化；以及10)ES细胞的心脏分化。课程将只使用在美国国立卫生研究院注册的人类胚胎干细胞系，特别是UC06，WA09和ES03。这门课程的一个主要长期目标将是分享和改进标准方案，并最终出版一本技术手册，在风格上类似于由Hogan等人编辑的流行的《操纵老鼠胚胎》手册。事实上，课程大纲目前包含了方案纲要、试剂清单和研究论文，为干细胞研究人员提供了宝贵的参考资源。这些材料可根据杰克逊实验室的要求获得。这些目标将通过向16名因其突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，紧接着将举行第二届国际干细胞倡议年度研讨会，来自世界各地主要ES细胞研究中心的人类ES细胞研究人员将在研讨会上讨论使用共享试剂和方法进行的实验的结果。课程结束后，还将在荒山岛生物实验室每年举办一次“陆地和海洋干细胞”研讨会，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无处不在的研究工具的主要瓶颈仍然是缺乏必要的动手培训来学习如何培养这些敏感细胞。本申请中建议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ES细胞的动手培训。

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.