Development of pig embryos after electro-activation and in vitro fertilization in PZM-3 or PZM supplemented with fetal bovine serum.
The aim of present study was to optimize culture conditions for pig embryos. Initially, we evaluated three different basic culture conditions. When embryos from electro-activation (parthenotes) or in vitro fertilization (IVF-embryos) were cultured in PZM supplemented with 3 mg/ml bovine serum albumin (PZM-3) in 4-well dishes, in medium covered with oil in 4-well dishes or in droplets under oil, 0%, 33% and 20% of the parthenotes, and 11%, 23% and 20% of the IVF-embryos developed to blastocysts. Subsequently, we examined the development of embryos when they were cultured in 4-well dishes in medium covered with oil continuously for 7 days or cultured under the same conditions but with a change to fresh medium on Days 2 and 4. In this experiment, 23% (no medium change) and 34% (change) of the parthenotes developed to blastocysts, respectively. When IVF-embryos were cultured under similar conditions, 33% and 38% of the embryos developed to blastocysts. Further improvement was achieved when PZM was supplemented with FBS from Day 4. In this experiment, 47% of the parthenotes developed to blastocysts with an average cell number of 57 +/- 7.7. In IVF-embryo group, 49% of the embryos developed to blastocysts with a mean cell number of 60 +/- 6.1. These results indicate that a change to fresh medium and inclusion of FBS in the medium during the late stages of culture can generate a higher proportion of high-quality blastocysts.
Okada
Konosuke
K
Center for Cell Therapy and Tissue Repair, VUZV Laboratory, Prague, Czech Republic. datti55@hotmail.com
Krylov
Vladimir
V
Kren
Radomir
R
Fulka
Josef
J
D000818:Animals; D001755:Blastocyst; D002417:Cattle; D003470:Culture Media; D004560:Electricity; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D005312:Fetal Blood; D009865:Oocytes; D013552:Swine; D015053:Zygote
D003470:Culture Media