Co-axial Microinjection System for Freezing and Biopsy of Early Embryos

用于早期胚胎冷冻和活检的同轴显微注射系统

• 批准号: 9031169
• 负责人: Paul Jon Taylor
• 金额: $ 59.5万
• 依托单位: GENESEARCH, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-05 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9031169
• 关键词: Address; Area; Arts; Biopsy; Bladder; Businesses; Bypass; Caliber; Cattle; Cell division; Cells; Certification; Chills; Collection; Communities; Contractor; Contracts; DNA; DNA analysis; Defect; Development; Devices; Electronics; Embryo; Embryo Research; Embryonic Development; Equipment; Equus caballus; Evaluation; Failure; Feedback; Fertility; Fiber Optics; Freezing; Funding; Genetic Materials; Glass; Goals; Health; Herpes zoster disease; Human; Injection of therapeutic agent; Inner Cell Mass; Instruction; Intellectual Property; Laboratories; Lasers; Legal patent; Licensing; Life; Light; Lighting; Liquid substance; Llama; Location; Mammals; Metals; Methods; Microinjections; Micromanipulation; Microscope; Modeling; Molds; Montana; Movement; Mus; Needles; Ovum; Penetration; Phase; Printing; Procedures; Process; Production; Puncture procedure; Regulatory Affairs; Research; Research Personnel; Resistance; Retrieval; Rupture; Sampling; Shapes; Shipping; Ships; Side; Special Equipment; Specialist; Staging; Stem cells; Sterilization; Stretching; Structure; Suction; Support System; Surface; System; Technology; Testing; Texas; Time; Trademark; Training; Transgenic Mice; Tube; United States National Institutes of Health; Universities; Uterus; Vacuum; Validation; Variant; Vision; Water; Work; Zona Pellucida; base; blastocyst; blastomere structure; capsule; cell water; clinical application; commercialization; design; experience; genetic disorder diagnosis; hatching; implantation; improved; preimplantation; prototype; seal; success; sucking; support network; systems research; tool; user-friendly

DESCRIPTION (provided by applicant): The Dracula Pipette is a co-axial embryo microinjection tool, with the injection pipette presented from inside the holding pipette. Double nested tips seal against the surface of the spherical embryo when a vacuum is applied in the space between them. This allows a portion of the nearside surface of an embryo to be held by vacuum while a glass pipette is passed for injection, aspiration of fluid contents or biopsy of embryonic cells. A proprietary laser-drilled biopsy probe, invented shortly before the beginning of the project, provides a new and better method for harm-free biopsy of pre-implantation embryos at a stage of development when they can provide more accurate DNA information for pre-implantation genetic diagnosis (PGD). The US Patent and Trademark Office issued the patent for the "suck-and-puncture" process in less than a year from the date of application, saying there was no relevant prior art to consider. Patents for the device itself and for our proprietary laser-drilled biopsy probe are pending. In October of 2011 this project was designated as high priority by NIH. This device, useful at all stages of embryo development up to the time of implantation in the uterus, is uniquely capable of control and manipulation of embryos at the hatched blastocyst stage of development. Hatching refers to the natural process of escape from the zona pellucida (the capsule that encloses the ovum and the embryo itself during the first few cell divisions). After hatching from the zona, these early embryos have no real structure and cannot be held on one side and penetrated from the opposite side, as is done routinely on embryos still in the zona, using conventional equipment. The Dracula Pipette has shown that it is valuable for water-volume reduction before freezing of embryos. Dr. Taylor, the Project Director, produced the first live babies from frozen/thawed embryos of llamas using this tool. Researchers at LSU, have recently had breakthrough success in freezing of large (900um) equine embryos with the Dracula system, which has not been possible before. Work done by Dr. Taylor at Montana State University with mouse embryos shows the capability of this system for survivable collection of a few stem cells from the inner cell mass and for injection of genetic material (stem cells) into the blastocoel space of mouse embryos for production of transgenic mice, a procedure that previously required special equipment for chilling of the zona, as well as the expensive piezo hammer for penetration of the zona. The human embryo may exist for more than two days as a hatched blastocyst, and there has been no good tool for manipulating it at this most-differentiated stage before implantation. Because it is large and fluid-filled, the human hatched blastocyst is difficult to freeze successfully and difficult to biopsy. Though the Dracula Pipette will surely be important for use with the expanded and hatched blastocysts of other species and on embryos at all stages before implantation, it is the only tool on the horizon for harm-free biopsy of human hatched blastocysts beyond the 8-cell stage and before implantation in the uterus. The Dracula Pipette Project will supply the current best prototype of the device for testing and practical use by several respected embryo researchers in a variety of laboratory settings, working mainly with bovine and mouse embryo models. The consultants will provide feedback about their experiences with the device and their ideas for improving or changing it for specific purposes. The inventor and a team of technical contractors will work to correct design problems as they are identified and will use appropriate micro-technology firms for production of specific improved prototype parts. As these cycles of evaluation and upgrade continue, emphasis will shift to practical manufacture of all parts, so a robust, user-friendly device can be presented to the scientific community. This tool, the Dracula Pipette, offers inexpensive embryo manipulation for routine procedures like water- volume reduction before freezing, and the capability to perform previously very difficult or impossible procedures on later-stage embryos. GeneSearch, Inc., the applicant Small Business, will commercialize the Dracula Pipette and biopsy probe through licensing of the intellectual property involved. We will maintain a small facility for creation and testing of new prototypes and variants of the device, and for coordinatin with licensees to provide instruction and practical assistance for new users.

描述（由申请人提供）：Dracula移液管是一种同轴胚胎微注射工具，其注射移液管从保持移液器内部显示。当在它们之间的空间中施加真空时，双嵌套的尖端密封在球形胚胎的表面上。这允许在传递玻璃移液器以进行注射，液体含量或胚胎细胞活检时，将胚胎的一部分近侧表面保持在真空中。项目开始前不久发明的专有激光钻孔的活检探针在开发阶段提供了一种新的，更好的方法，可以在开发阶段进行无害的植入前胚胎活检，以提供更准确的DNA信息，以提供更准确的DNA信息，以进行植入前植入前的遗传诊断（PGD）。美国专利商标办公室在不到一年的时间内就发布了“吮吸和功能”过程的专利，称没有相关的先前艺术需要考虑。该设备本身以及我们专有激光钻孔活检探针的专利正在待处理。 2011年10月，该项目被NIH指定为高度优先级。该设备在胚胎发育的所有阶段都有用，直到子宫内植入时期，在孵化的开发胚泡阶段都可以控制和操纵胚胎。孵化是指从Zona pellucida逃脱的自然过程（在前几个细胞分裂中封闭了卵子和胚胎本身的胶囊）。从Zona孵化后，这些早期的胚胎没有真实的结构，不能在一侧固定并从另一侧穿透，就像使用常规设备在Zona的胚胎上常规上一样。 Dracula移液器表明，在冷冻胚胎之前，它对于还原水量是有价值的。项目主管泰勒博士使用此工具从冰冻/融化的llamas胚胎中生产了第一批活婴儿。 LSU的研究人员最近在冻结大型（900UM）马胚胎的情况下取得了突破性的成功，而Dracula系统以前是不可能的。蒙大拿州立大学用小鼠胚胎的泰勒博士完成 材料（干细胞）进入小鼠胚胎的胚泡空间，用于生产转基因小鼠，该程序先前需要特殊设备来使Zona冷却，以及昂贵的压电锤以渗透Zona。作为孵化的胚泡可能存在人类的胚胎两天以上，并且在植入前最差异的阶段没有很好的工具来操纵它。因为它很大且充满液体，所以 孵化的胚泡很难成功冻结，难以活检。尽管植入前的其他物种的膨胀和孵化的胚泡肯定对于与其他物种的膨胀和孵化的胚泡一起使用肯定很重要，但它是植入前的所有阶段，但它是唯一的工具，即在8细胞阶段以及在子宫内植入之前对人类孵化的胚泡进行无害的活检。 Dracula移液器项目将为设备的当前最佳原型提供 几个受人尊敬的胚胎研究人员在各种实验室环境中的测试和实际使用，主要与牛和小鼠胚胎模型一起工作。顾问将提供有关他们在设备方面的经验以及为特定目的改进或更改其的想法的反馈。发明家和一组技术承包商将在确定的设计问题上纠正设计问题，并将使用适当的微技术公司来生产特定改进的原型零件。随着这些评估和升级周期的继续，重点将转移到所有零件的实际制造中，因此可以使用强大的，用户友好的设备 呈现给科学界。该工具是Dracula移液器，提供了廉价的胚胎操纵，以进行常规程序，例如冻结前的水量减少，以及在后期胚胎上执行以前非常困难或不可能的程序的能力。申请人小型企业Genesearch，Inc。将通过许可所涉及的知识产权的许可使Dracula移液器和活检调查商业化。我们将维护一个小型设施，用于创建和测试设备的新原型和变体，以及与被许可人的坐标素，以为新用户提供指导和实践帮助。