Growth factors effects on preimplantation development of mouse embryos exposed to tumor necrosis factor alpha.
The success rates of assisted reproduction techniques are still unsatisfactory. Relatively few in vitro cultured embryos reach the blastocyst stage. The purpose of the study was to evaluate the protective potential of epidermal growth factor (EGF), insulin-like growth factors 1 and 2 (IGF-I, IGF-II) and stem cell factor (SCF) on in vitro development of pre-implantation mouse embryos exposed to tumor necrosis factor alpha (TNFalpha). C3B6F1 female mice were superovulated with 5 IU of pregnant mare serum gonadotropin (PMSG) and 48 h later with 5IU of equine chorionic gonadotropin (eCG). Following the second injection females were mated with DBA males. Two cell embryos were flushed out from the fallopian tubes 40 h after eCG administration. After retrieval, the embryos were divided into control and experimental media and incubated in groups of ten for 96 h (37 degrees C, 5%CO(2), in droplets of 50 microl under mineral oil). In the first part of experiment, the embryo development was tested in media containing EGF, IGF-I, IGF-II, SCF, TNF-alpha (1 to 1000 ng/ml). In the second part of the study, the development of embryos was examined in medium containing 100 ng/ml TNFalpha and one of following factors: IGF-I, IGF-II; EGF or SCF (100 ng/ml). During the culture embryos were examined at 24 hours intervals to assess the embryo development. Blastocyst rate was determined following 96 hours of culture. Evaluation of total blastocyst cell number (TB) and inner cell mass (ICM) was also performed. TNFalpha significantly reduced (p<0.05) the blastocyst rates as well as TB and ICM. The examined growth factors improved the development of embryos exposed to TNFalpha. Thus, in this study, the protective action of IGF-I and II, EGF and SCF against the detrimental influence of TNFalpha was demonstrated.
Głabowski
Wojciech
W
Department Of Histology And Embryology, Pomeranian Medical University, Szczecin, Poland. wojtekgl@sci.pam.szczecin.pl
Kurzawa
Rafał
R
Wiszniewska
Barbara
B
Baczkowski
Tomasz
T
Marchlewicz
Mariola
M
Brelik
Paweł
P
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D002452:Cell Count; D006063:Chorionic Gonadotropin / Q000008:administration & dosage; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005260:Female; D006064:Gonadotropins, Equine / Q000008:administration & dosage; D006133:Growth Substances / Q000494:pharmacology*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D007335:Insulin-Like Growth Factor II / Q000494:pharmacology; D008297:Male; D051379:Mice; D008811:Mice, Inbred DBA; D011247:Pregnancy; D019089:Stem Cell Factor / Q000494:pharmacology; D013480:Superovulation; D020858:Tissue and Organ Harvesting; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D006063:Chorionic Gonadotropin; D006064:Gonadotropins, Equine; D006133:Growth Substances; D019089:Stem Cell Factor; D014409:Tumor Necrosis Factor-alpha; D004815:Epidermal Growth Factor; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II