Comparative fundamental cryobiology of mouse embryonic stem cells

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Comparative fundamental cryobiology of mouse embryonic stem cells

Please use this identifier to cite or link to this item: http://hdl.handle.net/10355/9881

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dc.contributor.advisor Critser, John Kenneth en_US
dc.contributor.author Benson, Corinna Mary Kashuba, 1971- en_US
dc.date.accessioned 2011-02-07T18:47:00Z
dc.date.available 2011-02-07T18:47:00Z
dc.date.issued 2009 en_US
dc.date.submitted 2009 Fall en_US
dc.identifier.other KashubaC-121409-D815 en_US
dc.identifier.uri http://hdl.handle.net/10355/9881
dc.description The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. en_US
dc.description Title from PDF of title page (University of Missouri--Columbia, viewed on January 25, 2011). en_US
dc.description Includes bibliographical references en_US
dc.description Vita. en_US
dc.description Thesis advisor: John K. Critser. en_US
dc.description "December 2009" en_US
dc.description Ph. D. University of Missouri-Columbia 2009. en_US
dc.description Dissertations, Academic -- University of Missouri--Columbia -- Veterinary pathobiology area program. en_US
dc.description.abstract Mouse embryonic stem cell (mESC) lines are central to projects such as the Knock-Out Mouse Project, which seek to create thousands of mutant mouse strains using mESCs for the production of human disease models. The ability to efficiently cryopreserve these cell lines for banking and transport is crucial to the success of these programs. The post-thaw recovery of viable cells varies significantly by genetic background, therefore there is a need to improve the efficiency and reduce the variability of current mESC cryopreservation methods. We employed the principles of fundamental cryobiology to improve the cryopreservation protocol of five mESC lines from different genetic backgrounds (BALB/c, C57BL/6, CBA, FVB, and 129R1 mESCs). Using methods outlined in this dissertation, a protocol utilizing 1 M propylene glycol, a cooling rate of 1°C/minute, and plunge into liquid nitrogen at -41°C, combined with subsequent warming in a 22°C water bath significantly improved post-thaw recovery for most mESC lines. Additionally, the effects of Latrunculin A (LATA), 1.5 M dimethyl sulfoxide (Me2SO), and temperature were examined on C57BL/6 mESC osmotic response and permeability parameters. Temperature, Me2SO, and LATA significantly influenced isosmotic cell volume, and LATA significantly affected adjusted osmotically inactive cell volume as well as permeability parameters for the C57BL/6 mESC line. en_US
dc.format.extent xi, 159 pages en_US
dc.language.iso en_US en_US
dc.publisher University of Missouri--Columbia en_US
dc.relation.ispartof 2009 Freely available dissertations (MU) en_US
dc.subject.lcsh Cryopreservation of organs, tissues, etc. -- Methods en_US
dc.subject.lcsh Cryobiology -- Methods en_US
dc.subject.lcsh Embryonic stem cells -- Cryopreservation en_US
dc.subject.lcsh Transgenic mice en_US
dc.subject.lcsh Glycols en_US
dc.subject.lcsh Thiazoles en_US
dc.subject.lcsh Dimethyl sulfoxide en_US
dc.title Comparative fundamental cryobiology of mouse embryonic stem cells en_US
dc.type Thesis en_US
thesis.degree.discipline Veterinary pathobiology area program en_US
thesis.degree.grantor University of Missouri--Columbia en_US
thesis.degree.name Ph. D. en_US
thesis.degree.level Doctoral en_US
dc.identifier.oclc 698474012 en_US
dc.relation.ispartofcommunity University of Missouri-Columbia. Graduate School. Theses and Dissertations. Dissertations. 2009 Dissertations


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