Effect of lipid polarization by centrifugation at different developmental stages on post-thaw survival of bovine in vitro produced 16-cell embryos.
The developmental rate to the blastocyst stage of frozen-thawed bovine in vitro produced embryos at stages earlier than Day 6 morula is not sufficiently high for practical utilization. The present study was undertaken to determine the effect of polarization of lipid droplets in the cytoplasm of bovine in vitro produced embryos from zygotes to the 8-cell stage, by centrifugation without following micromanipulation, on the survival rate of Day 4 16-cell embryos. After centrifugation at 15,500 x g in medium containing cytochalasin D, embryos were cultured to the 16-cell stage, classified as either mostly or partially delipidated by degree of lipid droplet removal, and then frozen. Embryos centrifuged at the 2-cell stage developed to the 16-cell stage similarly to those centrifuged at the 8-cell stage. The developmental rate to blastocysts after freezing of the mostly delipidated 16-cell embryos centrifuged at the 2-cell stage was higher than that of those centrifuged at the zygote stage, those that were partially delipidated at the 2-cell stage, and those that were not centrifuged. The results demonstrate that polarization of lipid droplets at the 2-cell stage by centrifugation without micromanipulation improved the survival rate of mostly delipidated 16-cell embryos after freezing.
Tominaga
K
K
Hyogo Prefectural Agricultural Institute, Kasai, Japan.
Shimizu
M
M
Ooyama
S
S
Izaike
Y
Y
D000818:Animals; D001391:Azo Compounds / Q000737:chemistry; D001562:Benzimidazoles / Q000737:chemistry; D002417:Cattle / Q000196:embryology* / Q000502:physiology*; D002498:Centrifugation / Q000662:veterinary; D004396:Coloring Agents / Q000737:chemistry; D015925:Cryopreservation / Q000662:veterinary; D015638:Cytochalasin D / Q000737:chemistry; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D005456:Fluorescent Dyes / Q000737:chemistry; D019787:Linoleic Acid / Q000737:chemistry; D008055:Lipids / Q000502:physiology*; D008297:Male; D008856:Microscopy, Fluorescence / Q000662:veterinary; D008858:Microscopy, Phase-Contrast / Q000662:veterinary; D009281:Naphthalenes; D019384:Nucleic Acid Synthesis Inhibitors / Q000737:chemistry; D011402:Pronase / Q000737:chemistry; D011419:Propidium / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D015053:Zygote / Q000166:cytology / Q000502:physiology
D001391:Azo Compounds; D001562:Benzimidazoles; D004396:Coloring Agents; D005456:Fluorescent Dyes; D008055:Lipids; D009281:Naphthalenes; D019384:Nucleic Acid Synthesis Inhibitors; D015638:Cytochalasin D; D012710:Serum Albumin, Bovine; D011419:Propidium; D019787:Linoleic Acid; C016118:Sudan Black B; D011402:Pronase; C017807:bisbenzimide ethoxide trihydrochloride