Biopsy and Freezing of Later-stage Mouse Blastocysts Using the Dracula Pipette

使用 Dracula 移液器对后期小鼠囊胚进行活检和冷冻

• 批准号: 8455935
• 负责人: EDWARD E SCHMIDT
• 金额: $ 10.69万
• 依托单位: GENESEARCH, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-19 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8455935
• 关键词: Address; Agreement; Aspirate substance; Biomedical Research; Biopsy; Businesses; Caliber; Cells; Chimera organism; Clinic; Contract Services; Cryopreservation; Culture Media; Development; Embryo; Embryology; Engineering; Equipment; Equus caballus; Fostering; Freezing; Future; Generations; Goals; Hand; Human; In Vitro; Individual; Injection of therapeutic agent; International; Laboratories; Lasers; Liquid substance; Llama; Maintenance; Marketing; Materials Testing; Methods; Molecular; Montana; Mus; Osmosis; Output; Penetration; Performance; Phase; Positioning Attribute; Procedures; Production; Property; Protocols documentation; Reproduction; Research; Side; Small Business Technology Transfer Research; Specialist; Staging; System; Technology; Testing; Time; Universities; Vacuum; Work; base; blastocyst; cost; design; embryo culture; embryonic stem cell; empowered; genetic analysis; hatching; improved; in utero; innovation; instrument; micromanipulator; novel; performance tests; preimplantation; pressure; prototype; public health relevance; seal; tool; user-friendly

DESCRIPTION (provided by applicant): Procedures on pre-implantation embryos are a cornerstone of much biomedical research; however in all cases, the procedures are very difficult and delicate, requiring exceptionally skilled technical experts and expensive equipment. The difficulties associated with these procedures often results in most such work being avoided by most biomedical research groups. When it must be performed, it is generally out-sourced to dedicated laboratories or facilities. Two downsides to this are: (1) many important questions are likely avoided due to the cost and difficulty of these procedures; and (2) many unique and potentially valuable mouse lines are likely lost or abandoned because the cost and/or complexity of cryopreservation are prohibitive. GeneSearch, Inc. has developed a novel co-axial embryo manipulation tool that promises to address these issues. The innovative elegance of this tool lies in its simple design, in which, rather than having separate "holding" and "manipulating" pipettes that work at opposite poles of the embryo, the Dracula has the manipulating pipette within the holding pipette. The major advantage is that the manipulating pipette contacts the embryo in a region that is constrained by the regulated forces imparted by the holding pipette. Original prototypes of the tool allowed, for the first time, dramatic advances in cryopreservation of large hatched embryos such as those of the llama and horse. Now, GeneSearch is scaling down the size of the tool, using new materials for some parts, and improving its handling specifically for manipulation of mouse-sized embryos. Phase I of the proposed study aims to optimize the "Mouse Dracula" tool for mouse embryological procedures including embryo cryopreservation, production of chimeric mice by ES cell injection, and survival-biopsy for genetic analyses. To accomplish this, we have proposed three specific aims, which (1) optimize tool performance, (2) develop improved protocols for mouse embryo cryopreservation, and (3) test the performance of the tool for chimera production and survival-biopsy-based genetic analyses. This is a collaborative research agreement between a Montana-based small business, GeneSearch Inc., and an embryologist/molecular geneticist at Montana State University. The goal of this Phase I STTR project is to prepare the Mouse Dracula tool to move on to Phase II market-oriented development within a one-year period.

描述（由申请人提供）：植入前胚胎的程序是许多生物医学研究的基石；但是，在所有情况下，这些程序都是非常困难和精致的，需要非常熟练的技术专家和昂贵的设备。与这些程序相关的困难通常会导致大多数生物医学研究小组避免了大多数这样的工作。当必须执行时，通常将其用于专用实验室或设施。这两个弊端是：（1）由于这些程序的成本和困难，可能避免了许多重要的问题； （2）由于冷冻保存的成本和/或复杂性是过于强烈的，许多独特且潜在的小鼠线可能会丢失或放弃。 Genesearch，Inc。开发了一种新颖的同轴胚胎操纵工具，该工具有望解决这些问题。该工具的创新优雅在于其简单的设计，在这种设计中，Dracula在胚胎相对的相对的两极中没有单独的“握住”和“操纵”移液器，而是在握持移液器中具有操纵移液管。主要优点是，操纵移液管接触的区域中，该区域受固定移液器施加的调节力的约束。该工具的原始原型首次允许急剧进步 在冷冻保存中，大型孵化的胚胎，例如骆驼和马的胚胎。现在，GeneSearch正在使用新材料来缩小工具的尺寸，并为某些零件进行新的材料，并专门改进其处理小鼠大小的胚胎。拟议研究的I期旨在优化用于小鼠胚胎学程序的“小鼠吸血鬼”工具，包括胚胎冷冻保存，通过ES细胞注射生产嵌合小鼠和用于遗传分析的生存期。为此，我们提出了三个特定目标，（1）优化工具性能，（2）为小鼠胚胎冷冻保存提供了改进的协议，以及（3）测试该工具的性能，用于嵌合体生产和基于生存的基因分析。这是蒙大拿州立大学的蒙大拿州小型企业Genesearch Inc.与胚胎学家/分子遗传学家之间的合作研究协议。 I阶段ISTTR项目的目标是准备一年内以II期市场为导向的开发的鼠标Dracula工具。