Current Protocols in Stem Cell Biology

描述(申请人提供)：我们目前正处于一场遗传学革命之中，这场革命有望将“突破性”生物医学研究推向新的水平。这场革命在很大程度上依赖于对基因之间的关系及其对复杂生物系统和表型的影响的分析。研究人员必须越来越熟练地操纵和使用遗传模型来解决各种重要的生物学问题。布什总统最近决定允许美国国立卫生研究院资助人类胚胎干细胞研究，这为美国调查人员创造了一个巨大的机会。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是学习如何培养这些敏感细胞所必需的动手培训。这项申请中提出的课程“干细胞生物学的现行方案”将为研究人员提供实践培训，以学习如何培养、操作和区分人类胚胎干细胞与体外培养的细胞。本课程将汇集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。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。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。这些目标将通过向26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。该课程将于2003年8月3日至8日在缅因州巴尔港的杰克逊实验室举行，随后将于8月9日至11日在荒漠山岛生物实验室举行“陆地和海洋干细胞”研讨会。

DESCRIPTION (provided by applicant): We are currently in the midst of a genetics revolution that promises to push "breakthrough" biomedical research to new levels. A large part of this revolution depends upon the analysis of the relationships between genes and their effects on complex biological systems and phenotypes. Investigators must become increasingly skilled in the manipulation and use of genetic models to address a variety of important biological questions. The recent decision by President Bush to allow National Institutes of Health funding of human embryonic stem (ES) cell research creates a tremendous opportunity for U.S. investigators. The current major bottleneck to human ES cells becoming a ubiquitous research tool in the biomedical research community is the hands-on training necessary to learn how to culture these sensitive cells. The course proposed in this application, "Current Protocols in Stem Cell Biology", will provide hands-on training for investigators to learn how to culture, manipulate, and differentiate human ES cells from in vitro. This course will bring together some of the leading experts on ES cell technology to train students in the successful culture, maintenance and manipulation of ES cells. A major long term goal for this course will be to share and improve standard protocols, and eventually to publish a technique manual, similar in style to the popular "Manipulating the Mouse Embryo" manual edited by Hogan et al. 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. These aims will be accomplished by offering an intensive 5-day course to 26 participants chosen for their outstanding research potential. The Course will be held annually in August 3-8, 2003 at The Jackson Laboratory in Bar Harbor, Maine, and will be followed by a companion symposium "Stem Cells on Land and at Sea" to be held at The Mount Desert Island Biological Laboratory on August 9-11.