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Title: BIRTH OF PIGLETS AFTER TRANSFER OF EMBRYOS CRYOPRESERVED BY CYTOSKELETAL STABILIZATION AND VITRIFICATION

Author
item Dobrinsky, John
item Pursel, Vernon
item LONG, C - 1265-10-00
item Johnson, Lawrence

Submitted to: Biology of Reproduction
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/30/1999
Publication Date: N/A
Citation: N/A

Interpretive Summary: Pig embryos suffer from severe sensitivity to hypothermic conditions. Most of the research has focused on their high lipid content, while little research has been conducted on structural damage to embryos during cryopreservation. Many cryoprotective agents function to depolymerize cytoskeletal components prior to cooling, however, some may be toxic to cells. The objectives of this study were to document cytoskeletal alterations during pig embryo vitrification, then inhibit disruptions during cryopreservation to determine the developmental competence of cytoskeletal stabilized and vitrified pig embryos. Vitrified pig embryos displayed microfilament disruptions and variable developmental competence. Treatments inhibited cytoskeletal disruptions and significantly improved survival of expanded and hatched blastocysts. After transfer of stabilized and vitrified/warmed embryos into surrogate females, two gilts farrowed live and normal offspring, with more pregnancies established. This study shows that cytoskeletal stabilization prior to vitrification improves blastocyst developmental competence after cryopreservation, producing piglets from embryos cryopreserved with vitrification. This technological advancement provides researchers and the swine industry for the first time a reliable method for the indefinite preservation of pig embryos.

Technical Abstract: Pig embryos suffer from severe sensitivity to hypothermic conditions. Most of the research has focused on their high lipid content, while little research has been conducted on structural damage to embryos during cryopreservation. Many cryoprotective agents function to depolymerize cytoskeletal components prior to cooling, however, some may be toxic to cells. The objectives of this study were to document microfilament alterations during swine embryo vitrification, utilize cytochalasin-b, a MF inhibitor, during cryopreservation to stabilize microfilaments, and to determine the developmental competence of cytoskeletal stabilized and vitrified pig embryos. Vitrified pig embryos displayed microfilament disruptions and variable developmental competence. Treatment with cytochalasin-b inhibited microfilament polymerization and significantly improved survival of expanded and hatched blastocysts. After transfer of stabilized and vitrified/warmed embryos into surrogate females, two gilts farrowed live and normal offspring, with more pregnancies established. This study shows that microfilament depolymerization prior to vitrification improves blastocyst developmental competence after cryopreservation, producing piglets from embryos cryopreserved with vitrification. This technological advancement provides researchers and the swine industry for the first time a reliable method for the indefinite preservation of pig embryos.