Effects of oviductal and cumulus cells on in vitro fertilization and embryo development of porcine oocytes fertilized with epididymal spermatozoa.
This study was designed to evaluate the effects of adding porcine oviductal epithelial cell (POEC) monolayers before or during the fertilization of denuded or cumulus-enclosed oocytes, in terms of fertilization results and subsequent embryo development. The variables determined were: penetration rate, mean number of spermatozoa per oocyte, male pronucleus formation rate, monospermy rate, cleavage rate after 48 h of fertilization, blastocyst rate, and mean number of nuclei per blastocyst. We used cumulus-free and cumulus-enclosed oocytes preincubated or fertilized in the presence of POEC, once the purity in epithelial cells of these cultures had been assessed. All the experiments involved the use of frozen-thawed epididymal spermatozoa to avoid replicate variability. The POEC cultures prepared showed a high proportion of epithelial cells (over 95%). Preincubation of oocytes with POEC before fertilization showed no effects on the fertilization variables determined. In contrast, during IVF under our experimental conditions, these cells attached to the cumulus cells and their interaction had a significant effect on some of the fertilization variables analyzed. The presence of POEC and cumulus cells during IVF increased oocyte penetrability. Moreover, in the absence of POEC, cumulus cells resulted in a reduced monospermy rate. On subsequent embryo culture, a lower cleavage and blastocyst formation rate were recorded when the oocytes had been preincubated with POEC before IVF.
Romar
R
R
Department of Physiology (Veterinary Physiology), Faculty of Veterinary Science, University of Murcia, 30071 Murcia, Spain. rromar@um.es
Coy
P
P
Ruiz
S
S
Gadea
J
J
Rath
D
D
D000818:Animals; D003470:Culture Media; D005314:Embryonic and Fetal Development; D004822:Epididymis / Q000166:cytology; D004847:Epithelial Cells; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D008297:Male; D009865:Oocytes / Q000502:physiology*; D006080:Ovarian Follicle / Q000166:cytology / Q000502:physiology; D010057:Oviducts / Q000166:cytology / Q000502:physiology; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000502:physiology*; D013552:Swine / Q000196:embryology / Q000502:physiology*
D003470:Culture Media