Combination effects of epidermal growth factor and glial cell line-derived neurotrophic factor on the in vitro developmental potential of porcine oocytes.
The developmental potential of in vitro matured porcine oocytes is still lower than that of oocytes matured and fertilized in vivo. Major problems that account for the lower efficiency of in vitro production include the improper nuclear and cytoplasmic maturation of oocytes. With the aim of improving this issue, the single and combined effects of epidermal growth factor (EGF) and glial cell line-derived neurotrophic factor (GDNF) on oocyte developmental competence were investigated. Porcine cumulus-oocyte cell complexes (COCs) were matured in serum-free medium supplemented with EGF (0, 10 or 50 ng/ml) and/or GDNF (0, 10 or 50 ng/ml) for 44 h, and subsequently subjected to fertilization and cultured for 7 days in vitro. The in vitro-formed blastocysts derived from selected growth factor groups (i.e. EGF = 50 ng/ml; GDNF = 50 ng/ml; EGF = 50 ng/ml + GDNF = 50 ng/ml) were also used for mRNA expression analysis, or were subjected to Hoechst staining. The results showed that the addition of EGF and/or GDNF during oocyte maturation dose dependently enhanced oocyte developmental competence. Compared with the embryos obtained from control or single growth factor-treated oocytes, treatment with the combination of EGF and GDNF was shown to significantly improve oocyte competence in terms of blastocyst formation, blastocyst cell number and blastocyst hatching rate (P < 0.05), and also simultaneously induced the expression of BCL-xL and TERT and suppressed the expression of caspase-3 in resulting blastocysts (P < 0.05). These results suggest that both GDNF and EGF may play an important role in the regulation of porcine in vitro oocyte maturation and the combination of these growth factors could promote oocyte competency and blastocyst quality.
Department of Animal Science,Faculty of Agriculture,University of Zabol,P. O. Box 98615-538,Zabol,Iran.
Mehdi Vafaye
MV
Valleh
Department of Basic Animal and Veterinary Sciences,Faculty of Life Sciences,University of Copenhagen,Copenhagen,Denmark.
Mikkel Aabech
MA
Rasmussen
Department of Basic Animal and Veterinary Sciences,Faculty of Life Sciences,University of Copenhagen,Copenhagen,Denmark.
Poul
P
Hyttel
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D053148:Caspase 3 / Q000235:genetics; D002478:Cells, Cultured; D054885:Cumulus Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005260:Female; D005306:Fertilization / Q000187:drug effects; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000494:pharmacology*; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods; D008297:Male; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D051020:bcl-X Protein / Q000235:genetics
D051100:Glial Cell Line-Derived Neurotrophic Factor; D018840:HSP70 Heat-Shock Proteins; D051020:bcl-X Protein; D004815:Epidermal Growth Factor; D053148:Caspase 3
Blastocyst quality; EGF; GDNF; In vitro maturation; Porcine