title: Birth of piglets after transfer of embryos cryopreserved by cytoskeletal stabilization and vitrification.
abstract: Pig embryos suffer severe sensitivity to hypothermic conditions, which limits their ability to withstand conventional cryopreservation. Research has focused on high lipid content of pig embryos and its role in hypothermic sensitivity, while little research has been conducted on structural damage. Documenting cytoskeletal disruption provides information on embryonic sensitivity and cellular response to cryopreservation. The objectives of this study were to document microfilament (MF) alterations during swine embryo vitrification, to utilize an MF inhibitor during cryopreservation to stabilize MF, and to determine the developmental competence of cytoskeletal-stabilized and vitrified pig embryos. Vitrified morulae/early blastocysts displayed MF disruptions and lacked developmental competence after cryopreservation; hatched blastocysts displayed variable MF disruption and developmental competence. Cytochalasin-b did not improve morula/early blastocyst viability after vitrification; however, it significantly (P < 0.05) improved survival and development of expanded and hatched blastocysts. After embryo transfer, we achieved pregnancy rates of almost 60%, and litter sizes improved from 5 to 7.25 piglets per litter. This study shows that the pig embryo cytoskeleton can be affected by vitrification and that MF depolymerization prior to vitrification improves blastocyst developmental competence after cryopreservation. After transfer, vitrified embryos can produce live, healthy piglets that grow normally and when mature are of excellent fecundity.
authors.affiliation: Germplasm & Gamete Physiology Laboratory and Gene Evaluation & Mapping Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Beltsville, Maryland 20705, USA. bigjohn@lpsi.barc.usda.gov
authors.forename: J R
authors.initials: JR
authors.lastname: Dobrinsky
authors.forename: V G
authors.initials: VG
authors.lastname: Pursel
authors.forename: C R
authors.initials: CR
authors.lastname: Long
authors.forename: L A
authors.initials: LA
authors.lastname: Johnson
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure; D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000276:immunology; D002462:Cell Membrane / Q000187:drug effects / Q000648:ultrastructure; D015925:Cryopreservation / Q000379:methods*; D003571:Cytochalasin B / Q000494:pharmacology; D003599:Cytoskeleton / Q000648:ultrastructure*; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000166:cytology* / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro; D008098:Litter Size; D011247:Pregnancy; D018873:Pregnancy Rate; D013552:Swine
chemical_list: D003571:Cytochalasin B