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（申请人提供）：德古拉移液器是一种同轴胚胎显微注射工具，注射吸管从持有移液器内部呈现。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的表面上。这允许在真空中保持胚胎近侧表面的一部分，同时通过玻璃移液管进行注射，液体内容物的抽吸或胚胎细胞的活检。在项目开始前不久发明的一种专有的激光钻孔活检探针，为胚胎发育阶段的植入前胚胎提供了一种新的、更好的无害化活检方法，可以为植入前遗传学诊断（PGD）提供更准确的DNA信息。美国专利商标局（US Patent and Trademark Office）在申请之日起不到一年的时间里就授予了“吸穿法”专利，称没有相关的现有技术需要考虑。该设备本身和我们专有的激光钻孔活检探针的专利正在申请中。2011年10月，该项目被NIH列为高优先级项目。该装置适用于胚胎发育的所有阶段，直到植入子宫的时间，具有独特的控制和操纵囊胚发育阶段的胚胎的能力。孵化是指从透明带（在最初的几次细胞分裂中包裹卵子和胚胎本身的荚膜）中逃脱的自然过程。从卵带中孵化出来后，这些早期胚胎没有真正的结构，不能像在仍在卵带中的胚胎上使用常规设备所做的那样，从一侧握住并从另一侧穿透。德古拉移液管已经证明了它在胚胎冷冻前减少水量的价值。项目主任泰勒博士使用该工具从冷冻/解冻的大羊驼胚胎中生产了第一批活的婴儿。路易斯安那州立大学的研究人员最近在用德古拉系统冷冻大型（900毫米）马胚胎方面取得了突破性的成功，这在以前是不可能的。蒙大拿州立大学的泰勒博士对小鼠胚胎所做的工作表明，该系统能够从内部细胞群中收集到一些存活的干细胞，并能够注射基因