Low concentrations of MEM vitamins during in vitro maturation of porcine oocytes improves subsequent parthenogenetic development.
To investigate the effects of water-soluble vitamin supplementation for IVM/IVC of porcine oocytes and evaluate maturation and developmental capacity in vitro, porcine cumulus oocyte complexes (COCs) was matured in NCSU-23-based medium with water-soluble vitamins for 44 h and then cultured in PZM-3 for 7 days following activation. The COCs were allocated into five treatment groups and matured in various concentrations of MEM vitamins (control, 0.05, 0.1, 0.2, 0.4, and 1x). Metaphase II plates of the cumulus-free oocytes were observed following Hoechest 33258 staining. The COCs were allocated into four treatment groups, matured in various concentrations of MEM vitamins (control, 0.05, 0.1, 0.2, and 0.4x) and cultured in PZM-3 following activation. Also, COCS were matured without MEM vitamins and cultured in PZM-3 with various concentrations (control, 0.1, 0.4, 1.0, and 2.0 x) of MEM vitamins. Furthermore, 2 x 2 factorial (IVM/IVC) experiments were performed in IVM medium with or without 0.05 x MEM vitamins and IVC medium with or without 0.4x MEM vitamins to examine the in vitro development of parthenogenetic embryos. Maturation rates of COCs treated with MEM vitamins did not differ significantly among groups. However, compared to the control group, oocytes matured with the addition of 0.05 x MEM vitamins developed to blastocysts at a higher percentage (P<0.05) following activation and culture in PZM-3 without MEM vitamins. Total cell number of blastocysts was significantly higher in the 0.05 x group. Addition of 0.4x MEM vitamins decreased (P<0.05) cleavage and blastocyst developmental rates compared with 0.05 x MEM vitamins-treated group. In contrast, addition of vitamins to PZM-3 medium for in vitro culture of activated porcine oocytes did not affect development. In conclusion, addition of a low concentration of MEM vitamins to IVM medium for porcine oocytes enhanced subsequent development and improved embryo quality.
Naruse
Kenji
K
Division of Animal Science and Resources, Research Center for Transgenic Cloned Pigs, Chungnam National University, Daejeon City 338-708, South Korea.
Kim
Hong Rye
HR
Shin
Young Min
YM
Chang
Suk Min
SM
Lee
Hye Ran
HR
Park
Chang Sik
CS
Jin
Dong Il
DI
D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000662:veterinary; D003470:Culture Media; D004305:Dose-Response Relationship, Drug; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D008677:Metaphase / Q000502:physiology; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology*; D010312:Parthenogenesis; D011897:Random Allocation; D013552:Swine / Q000196:embryology / Q000502:physiology*; D013997:Time Factors; D014815:Vitamins / Q000494:pharmacology*
D003470:Culture Media; D014815:Vitamins