Effect of protein supplement source on porcine oocyte maturation and subsequent embryonic development after parthenogenetic activation.
The aim of this study was to compare the effect of purified GPBoS and commonly used FCS on porcine oocyte maturation and subsequent embryonic development after their parthenogenetic activation. COCs were obtained from dissected follicles and cultured for 18, 24, 30, 36, 42 and 48 h in M-199 medium either with GPBoS or FCS. After 24 h with GPBoS, 91% of oocytes reached MI stage while in the medium supplemented with FCS, only 29% of oocytes reached the same stage (P < 0.05). The majority of oocytes from the FCS group (61%) reached MI stage approximately 6 h later. In the time periods between 36 to 48 h both groups of oocytes reached the same stage of maturation. After 48 h of culture the oocytes with extruded polar bodies were activated by a single electric pulse and then cultured with 4 mM 6-DMAP. Activated oocytes were cultured in PZM-3 medium supplemented with 3 mg/ml of BSA. After 7 days, the development and the quality of embryos were evaluated. The results showed that the maturation of oocytes in the presence of GPBoS significantly increased their subsequent developmental ability when compared with FCS supplementation (27% vs. 19% of blastocysts, P < 0.05). However, differential staining revealed that once blastocysts were formed in either group, they had the same total cell number (40 vs. 41) and also the ICM/total cell ratio (0.27 vs. 0.29).
Krylov
V
V
Center for Cell Therapy and Tissue Repair, VUZV Labs., Prague, Czech Republic. vkrylov@natur.cuni.cz
Kren
R
R
Okada
K
K
Vacková
I
I
Tlapáková
T
T
Fulka
J
J
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D001798:Blood Proteins / Q000378:metabolism / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D049108:Cell Enlargement / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000378:metabolism / Q000494:pharmacology*; D046149:Embryo Culture Techniques / Q000379:methods*; D005260:Female; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D034421:Sus scrofa; D013997:Time Factors
D001798:Blood Proteins; D003470:Culture Media