Utility of rapidly matured oocytes as recipients for production of cloned embryos from somatic cells in the pig.
The present study was conducted to examine the utility of rapidly matured oocytes as recipients for production of porcine embryos reconstituted with adult skin fibroblasts and whether arrest of meiotic resumption of recipient oocytes at the germinal vesicle (GV) stage by dibutyryl cyclic AMP (dbcAMP) improves in vitro developmental rates after reconstruction. At 24 h of maturation in the medium, 36.3% of oocytes reached the metaphase II (MII) stage. At 30 h of maturation, the percentage (71.4%) of MII oocytes did not significantly differ from that (78.0%) at 42 h of maturation. When MII oocytes recovered at 24 h of maturation were used as recipients, 22/156 (14.1%) cloned embryos developing to the blastocyst stage was significantly (P < 0.05) higher than those of embryos reconstituted with oocytes collected at 30 h (5/168; 3.0%) and 42 h (13/217; 6.0%) of maturation. Culture of oocytes in medium containing 1 mM dbcAMP for 20 h maintained 72.9% in the GV stage, whereas only 15.0% of nontreated oocytes were in the GV stage (P < 0.05). The effect of dbcAMP was reversible. However, the treatment of recipient oocytes with dbcAMP did not affect the development of reconstructed embryos when compared with nontreated oocytes. These results indicate that rapidly matured oocytes are superior in their ability to support development of porcine reconstructed embryos; however, arrest of meiotic resumption of recipient oocytes at the GV stage by dbcAMP does not improve reconstructed embryo developmental rates.
Department of Animal and Dairy Science, University of Georgia, 425 River Road, Athens, GA 30602-2771, USA.
Kazuchika
K
Miyoshi
S Jacek
SJ
Rzucidlo
Scott L
SL
Pratt
Steven L
SL
Stice
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D019976:Cloning, Organism / Q000379:methods*; D004431:Ear, External / Q000166:cytology / Q000502:physiology; D004624:Embryo Transfer; D005260:Female; D005347:Fibroblasts / Q000502:physiology; D018858:Germinal Center / Q000502:physiology; D008677:Metaphase / Q000502:physiology; D009865:Oocytes / Q000502:physiology*; D011247:Pregnancy; D013552:Swine
D003994:Bucladesine