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Mouse Stem Cell Research Using the Dracula Pipette

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 1R42OD018404-01A1
Agency Tracking Number: R42OD018404
Amount: $149,995.00
Phase: Phase I
Program: STTR
Solicitation Topic Code: 100
Solicitation Number: PA13-224
Solicitation Year: 2016
Award Year: 2015
Award Start Date (Proposal Award Date): 2015-03-15
Award End Date (Contract End Date): 2017-03-14
Small Business Information
676 S. Fergusen Ave., Suite 7
Bozeman, MT 59718-1951
United States
DUNS: 962692997
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 (406) 994-6375
Business Contact
Phone: (406) 686-4723
Research Institution
309 Montana Hall
Bozeman, MT 59717
United States

 Nonprofit college or university

DESCRIPTION provided by applicant Advanced approaches for genetic or epigenetic modification of embryonic stem ES cells including TALEN ZFN or CRISPR catalyzed genetic modifications or modulation of gene expression via small RNAs synthetic mRNAs or recombinant transcription factors have revolutionized our ability to modify the cells from which novel mouse lines are produced In contrast procedures for utilizing these cells within the context of a recipient embryo have changed little in decades Most embryological procedures require skilled expertise to perform some e g those involving hatched or manipulated blastocysts are inaccessible to even expert embryologists We have developed a novel co axial embryo manipulation tool the andquot Mouse Dracula Pipetteandquot which addresses these issues This innovative tool is based on an elegant patented concept in which rather than having separate andquot holdingandquot and andquot manipulatingandquot pipettes that work at opposite poles of the embryo the manipulating pipette is contained within the holding pipette The major advantage is that the manipulating pipette contacts the embryo in a region that is constrained under the regulated forces imparted by the holding pipette The original Dracula Pipette was conceived and developed by GeneSearch Inc for use with llama blastocysts which are mm in diameter and had been impossible to cryopreserve By facilitating replacement of blastocoel fluid with cryoprotectant and flushing of this at thawing this tool allowed the first reliable freezing and post thaw reanimation of llama embryos Reflecting the size of llama blastocysts the original Dracula was large hand held and constructed from drawn glass More recently we have developed the andquot Mouse Draculaandquot for small embryos like those of mice or humans which are times smaller in volume than llama blastocysts Scaling down to this size required high performance materials precision machining and micromanipulator control The tool was optimized for procedures including general manipulations cryopreservation cell injection and biopsy of trophectoderm cells of mouse blastocysts Advanced materials meld performance with glass like transparency Precision linear motion of the injection biopsy probe is hydraulic using a metal bellows The fine control afforded by the Mouse Dracula makes it effective for harm free manipulations even on delicate hatched blastocysts Here we propose a Fast Track application that will refine the Mouse Dracula for stem cell procedures and aim toward commercialization for these applications In Phase of this Fast Track application we propose three Specific Aims In Ph Aim we will optimize use of the Mouse Dracula to inject ES cells into the blastocoel for production of chimeric mice In Ph Aim we will optimize the tool for harm free biopsy of cells from the inner cell mass ICM In each case we will test early expanded and hatched blastocysts Evaluation will include assessment of the quality of pups born from the manipulated embryos as well as the reliability of biopsied materials for transfer into recipient embryos ex vivo culture or genetic analyses Achievement of these performance milestones will mark completion of Phase and initiate commencement of Phase In Phase we propose two Specific Aims In Ph Aim we will further refine andquot ease of useandquot and platform adaptability of the Mouse Dracula to make it optimally convenient for integration into current mouse embryology facilities In Ph Aim we will further refine the Mouse Dracula within the confines of the performance criteria achieved in the other aims for more efficient manufacture and more effective commercialization Thus we will investigate means of more efficient high throughput out sourcing of component production and assembly and investigate overall manufacturing marketing and distribution strategies for delivering the Dracula technology at a favorable cost to mouse embryology laboratories of all sizes RELEVANCE Currently mouse embryological procedures are accessible to only highly skilled technical experts Also some embryological stages e g hatched blastocysts or some procedures e g biopsy of cells from the ICM are so delicate that even in skilled hands they are not accessible Phase of this proposal optimizes a tool for mouse stem cell associated embryo manipulations that will make all procedures more accessible to a broader user group and will facilitate some currently refractory procedures Phase aims toward optimization of the user interface and improvements in manufacture that will favor commercialization of this tool for biomedical research applications

PUBLIC HEALTH RELEVANCE While technologies for modifying stem cells in culture have advanced in recent years technologies for using these cells in recipient embryos have not advanced substantially Existing methods are technically challenging cannot effectively achieve reliable harm free stem cell biopsy and are not practical for use on late stage hatched blastocysts Thus many embryological technologies are out of reach for most small or unspecialized biomedical research labs and some important conceivable manipulations are out of reach for all laboratories GeneSearch Inc previously developed a novel embryo manipulation tool the andquot Dracula Pipetteandquot for cryopreservation of llama embryos and patented the innovative coaxial design of the Dracula technology We recently developed an advanced version of the Dracula optimized for use with the much smaller embryos of mice or humans The andquot Mouse Draculaandquot will provide an affordable user friendly system that will empower a much larger number of embryology or reproduction labs to generate chimeric mice or recover stem cells from early or late stage blastocysts In addition we anticipate that the delicate manipulations afforded by this tool will empower new manipulations that cannot be reasonably achieved by contemporary approaches In addition to optimizing this tool and protocols using it for stem cell procedures on early and late stage mouse blastocysts within Phase this Fast track proposal includes Phase technological refinements aimed at optimizing the user interface and better positioning the tool for future commercial production and a Commercialization Plan that extends beyond this

* Information listed above is at the time of submission. *

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