Ultra-structural changes and developmental potential of porcine oocytes following vitrification.
This study evaluated the effects of exposure and/or vitrification of porcine metaphase II (MII) oocytes on their in vitro viability and ultra-structural changes with two experiments. Experiment 1 examined the effect of vitrified oocytes on microtubule localization, mitochondrial morphology, chromosome organization and the developmental rate in IVF control and vitrified oocytes. Oocytes matured for 44 h were subjected to IVF (IVF control). Oocytes matured for 42 h were exposed to cryoprotectants (CPA control), followed by 2h culture, and subjected to IVF. Oocytes vitrified at 42 h post-maturation were warmed, cultured for 2h, and subjected to IVF (vitrified). Experiment 2 evaluated the effect of oocytes freezing on development of ICSI with and without activation and parthenotes. Fresh and vitrified oocytes were subjected to ICSI with and without electrical activation. Cleavage and blastocyst rates were significantly (P<0.05) lower in vitrified IVF, parthenote and ICSI embryos than those in fresh counterparts. Between ICSI embryos from fresh oocytes and vitrified oocytes, the rates of blastocyst were significantly higher (P<0.05) in activated group than the group without activation. Significant differences (P<0.05) were observed in normal spindle configuration of vitrified (43.5%) compared to control (81.0%) oocytes, but no significant difference was observed between CPA exposed and control groups. In conclusion, porcine oocytes at MII stage are very sensitive to vitrification with altered microtubule localization and mitochondrial organization thus resulting in impaired fertilization and embryo development.
Shi
Lian-Yu
LY
College of Veterinary Medicine, Gyeongsang National University, Chinju 660-701, Republic of Korea.
Jin
Hai-Feng
HF
Kim
Jung-Gon
JG
Mohana Kumar
B
B
Balasubramanian
S
S
Choe
Sang-Young
SY
Rho
Gyu-Jin
GJ
D000818:Animals; D002875:Chromosomes / Q000502:physiology; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D046149:Embryo Culture Techniques / Q000662:veterinary*; D005307:Fertilization in Vitro / Q000662:veterinary*; D008677:Metaphase; D008928:Mitochondria / Q000502:physiology; D009865:Oocytes / Q000166:cytology / Q000254:growth & development* / Q000648:ultrastructure*; D020554:Sperm Injections, Intracytoplasmic / Q000662:veterinary; D013552:Swine