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An Intersection Of Science & Art: Vitrification Approaches And Open-Fabricated Tools For The Biomedical Model Sea Hare, Aplysia Californica, Allyssa M. Oune
An Intersection Of Science & Art: Vitrification Approaches And Open-Fabricated Tools For The Biomedical Model Sea Hare, Aplysia Californica, Allyssa M. Oune
LSU Master's Theses
The California sea hare (Aplysia californica) is an important biomedical model for molecular neurobiology, electrophysiology, learning, and memory due to their well-mapped and large neurons and well-characterized learning capabilities. The National Resource for Aplysia (NRA, University of Miami) maintains large stocks of live animals and relies on regular shipments of wild-caught individuals to maintain genetic diversity. This is labor and cost-intensive, and environmental changes could alter the availability of wild animals increasing the need to preserve this genetic resource. One solution is vitrification, ultra-fast cooling which produces an amorphous glass that minimizes damage to cells. Aplysia californica presents …
Effects Of Microtubule Stabilizing And Recovery Agents On Vitrified Bovine Oocytes, Emily Dzekunskas
Effects Of Microtubule Stabilizing And Recovery Agents On Vitrified Bovine Oocytes, Emily Dzekunskas
LSU Master's Theses
Oocyte vitrification, while beneficial for research and species conservation, has limited success due to cryoinjury to the meiotic spindle. This study aimed to evaluate treatments to improve meiotic spindle recovery after vitrification by treating bovine oocytes with either a microtubule stabilizing agent, or a microtubule recovery agent. In the first two experiments, Taxol and epothilone B were used to treat bovine oocytes before vitrification. Both compounds have microtubule stabilizing properties and are known antimitotic compounds commonly used in cancer treatment. Oocytes were incubated with one of the microtubule stabilizing agents for 15 minutes before vitrification at one of three concentrations …
Effects Of Dimethyl Sulfoxide And Glycerol Based Vitrification Protocols On Zona Pellucida Hardening In Mature Bovine Oocytes, Kaci Denise Rogers
Effects Of Dimethyl Sulfoxide And Glycerol Based Vitrification Protocols On Zona Pellucida Hardening In Mature Bovine Oocytes, Kaci Denise Rogers
LSU Master's Theses
Zona pellucida hardening is a natural process that occurs after oocyte fertilization to prevent polyspermic fertilization and to protect embryonic development. Pre-fertilization hardening of the zona pellucida however, decreases fertilization rates. Cryoprotectants have also been shown to negatively affect fertilization rates, one possible mechanism of which being through zona hardening. This experiment was conducted to determine the effect of different cryoprotectants on hardening of the zona pellucida using fresh mature bovine oocytes and vitrified mature bovine oocytes. Oocytes were post-slaughter from mixed breed cows. After collection, oocytes were randomly assigned to three cryoprotectant treatment groups: dimethyl sulfoxide (DMSO), glycerol or …
Vitrification Of Immature And Mature Bovine Oocytes, Paige T. Hardin
Vitrification Of Immature And Mature Bovine Oocytes, Paige T. Hardin
LSU Master's Theses
Vitrification is the latest technique used in cryopreservation, the ability to utilize this method with oocytes and embryos has become a valuable system. Vitrification has been successful with bovine embryos and oocytes but is far from optimal. Following cryopreservation storage discarding embryos can cause ethical issues, and mature oocytes have fragile organelles that can be detrimentally affected by ice crystal formation during freezing. Immature oocytes have not formed some of these temperature sensitive microfilaments and can circumvent these detrimental effects. The common intracellular cryoprotective agents are dimethyl sulfoxide, glycerol and ethylene glycol. Different combination of these agents have been reported …
Vitrification Of Equine Expanded Blastocysts, Fabian Andres Diaz
Vitrification Of Equine Expanded Blastocysts, Fabian Andres Diaz
LSU Master's Theses
The cryopreservation of equine expanded blastocysts (>300 um) has been largely unsuccessful primarily due to the low permeability to cryoprotectants and the large size of the equine embryo. Mechanical alternatives may provide means to overcome the capsule barrier and the relative large embryo size. In this regard, multiple experiments were performed in this study to evaluate different approaches of capsule puncture and blastocoele fluid extraction with the objective to develop a cryopreservation protocol for Day 8 equine expanded blastocysts. In the first experiment, twenty-four Day 8 expanded blastocysts were exposed to standard equine embryo vitrification solutions following one- or …
Cryopreservation Of Bovine And Caprine Oocytes By Vitrification, Sabrina Marie Luster
Cryopreservation Of Bovine And Caprine Oocytes By Vitrification, Sabrina Marie Luster
LSU Master's Theses
Cryopreservation of animal oocytes will permit germplasm of valuable or unique females to be preserved for extended times. The objective of this research was to derive a procedure to cryopreserve bovine oocytes by vitrification to be used as recipients for somatic cell nuclear transfer (SCNT). Caprine oocytes vitrified by the same procedure were assayed by cytological examination of microtubules. In the first two of three experiments, bovine oocytes matured in vitro were vitrified in a mixture of ethylene glycol (EG), dimethylsulfoxide (Me2SO) and trehalose, and then subjected to in vitro fertilization (IVF) or SCNT. For vitrification, oocytes were first exposed …