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

Genesearch, Inc., Bozeman MT

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Abstract

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.