Skip to main content
ARS Home » Research » Publications at this Location » Publication #85462
Title: CRYOPRESERVATION OF SWINE EMBRYOS: IN VITRO AND IN VIVO DEVELOPMENTAL COMPETENCE

Author
item Dobrinsky, John
item Pursel, Vernon
item Long, Charles
item Johnson, Lawrence

Submitted to: Journal of Animal Science
Publication Type: Abstract Only
Publication Acceptance Date: 3/15/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Pig embryos are sensitive to chilling, making them difficult to cryopreserve. This is less apparent in embryos of most mammalian species and is linked to the high lipid content and fragile cytoskeleton of pig embryos. Many cryoprotective agents act to depolymerize cytoskeletal components prior to cooling although this may be toxic to cells. We documented microfilament (MF) disruption during vitrification and utilized a MF inhibitor, cytochalasin-b (cyto-b) to stabilize MF. Laser scanning confocal microscopy revealed reduced fluorescence intensity of MF in cyto-b treated embryos, indicating successful MF depolymerization prior to cryopreservation. Morulae and early blastocysts (MB; n=34), expanded blastocysts (XB; n=52) and hatched blastocysts (HB; n=120) were vitrified with or without cyto-b. While MB did not survive cryopreservation (0%), treatment with cyto-b did not improve their viability (6%, p>0.05). However, cyto-b significantly improved XB (22 vs 60%; p<0.01) and HB (28 v 90%; p<0.01) development in vitro. Cyto-b treated and vitrified HB were transferred into two recipient females after warming. Four live and developmentally normal fetuses were recovered at 25 days of gestation from one recipient. These experiments show that the cytoskeleton is affected during vitrification, and that MF depolymerization prior to cryopreservation improves embryonic development.