Effect of culture medium and prostaglandin I(2) (PGI(2)) analogue on in vitro development of parthenogenetically activated cat oocytes.
A method of reliably producing developmentally competent cat embryos in vitro is a prerequisite for study of the physiology of early development and application of assisted reproductive techniques. Oocytes were collected and then cultured in TCM-199 + 10% FBS for 4 h. The matured oocytes were activated with a 20 microsec electric pulse at 1.2 kV/mm. The activated oocytes were incubated in 2 mM of 6-dimethylaminopurine (6-DMAP) for 4 h and were then divided randomly among the treatment groups. In experiment 1, we compared the effects of three culture systems (TCM-199, CR1-aa and Tyrode's) on the in vitro development of parthenogenetically activated cat oocytes. In experiment 2, we investigated the effect of addition of Iloprost (a stable prostaglandin I(2) analogue) to Tyrode's medium on in vitro development of parthenogenetically activated oocytes. As a control, we recovered in vivo produced blastocysts and determined their average cell number. In experiment 1, the cleavage frequency of the oocytes cultured in TCM199, CR1-aa and Tyrode's media were similar (74, 72 and 83%, respectively). However, the incidence of in vitro development to the blastocyst stage was significantly higher in Tyrode's medium (20.4%) than in TCM-199 (2.4%) or CR1-aa (11.1%). Likewise, the average cell number of in vitro activated blastocysts was higher in Tyrode's than in CR1-aa or TCM-199 (106.5 +/- 45.2 vs. 68.3 +/- 25.4 and 35.0 +/- 7.7, respectively; P<0.05). In experiment 2, the percentage of parthenogenetically activated oocytes that underwent in vitro blastocyst development was significantly improved by addition of Iloprost to the culture medium (33.6 vs. 19.1%; P<0.05). The average cell number of in vivo blastocysts (909.0 +/- 226.4) was significantly higher than those of in vitro blastocysts cultured in Tyrode's medium supplemented with or without Iloprost (103.2 +/- 31.3 and 112.2 +/- 39.3, respectively; P<0.05). This result indicated that the current culture method for cat pathogenetically activated oocytes requires further improvement.
Yin
Xi-Jun
XJ
Department of Animal Science and Technology, Sunchon National University, Suncheon, Korea.
Lee
Hyo-Sang
HS
Jeon
Jin-Tae
JT
Kong
Il-Keun
IK
D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002415:Cats; D002452:Cell Count; D003470:Culture Media / Q000494:pharmacology*; D046149:Embryo Culture Techniques; D005260:Female; D016285:Iloprost / Q000494:pharmacology*; D009865:Oocytes / Q000187:drug effects* / Q000254:growth & development*; D010312:Parthenogenesis
D003470:Culture Media; D016285:Iloprost