Effects of epidermal growth factor on early embryonic development after in vitro fertilization of oocytes collected from ewes treated with follicle stimulating hormone.
Epidermal growth factor (EGF) has been shown to enhance the in vitro rate of blastocyst formation in several species. Follicular development was induced in ewes (n=15) by twice daily administration of FSH-P on Days 13 and 14 of the estrous cycle. Cumulus oocyte complexes (COCs) were collected from all visible follicles (n=25+/-2.4/ewe) on Day 15. COCs from each ewe were cultured separately for 24h in maturation medium (containing 10% serum, LH, FSH and estradiol) with (8.2+/-0.9 per ewe) or without (7.8+/-0.8 per ewe) EGF (10 ng/ml). Oocytes were then denuded by hyaluronidase treatment, and healthy oocytes were cultured in the presence of frozen-thawed semen in synthetic oviductal fluid (SOF) medium containing 2% sheep serum. After 18-20 h, zygotes were transferred to SOF medium without glucose and cultured for about 36 h until they reached the 4-8 cell stage. Embryos were transferred to SOF medium with glucose for further development. Medium was changed every other day until blastocyst formation on Day 8 of culture (Day 1=day of fertilization). The rate of embryonic development was evaluated throughout the culture period. After maturation, cumulus cells were more expanded in the presence than in the absence of EGF. The rates of fertilization (overall 75.7+/-3.9%) and morula formation (overall 40.6+/-7.1%) were similar (P>0.05) for COCs cultured with or without EGF. However, EGF increased (P<0.01) the number of blastocysts (1.4+/-0.1 versus 0.6+/-0.2 per ewe) and tended to increase (P<0.1) the rate of blastocyst formation (21.0+/-6.6% versus 13.4+/-4.3% per ewe). These data demonstrate that EGF increases blastocyst formation in FSH-treated ewes. Therefore, EGF is recommended as a supplement to maturation medium to enhance embryonic development in vitro in FSH-treated sheep.
Grazul-Bilska
A T
AT
Department of Animal and Range Sciences, North Dakota State University, Fargo, ND, USA. anna-grazul-bilska@ndsu.nodak.edu
Choi
J T
JT
Bilski
J J
JJ
Weigl
R M
RM
Kirsch
J D
JD
Kraft
K C
KC
Reynolds
L P
LP
Redmer
D A
DA
D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000502:physiology; D005314:Embryonic and Fetal Development / Q000187:drug effects* / Q000502:physiology; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D008297:Male; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology*; D011247:Pregnancy; D012756:Sheep / Q000196:embryology / Q000502:physiology*
D004815:Epidermal Growth Factor; D005640:Follicle Stimulating Hormone