Effect of co-culture of porcine sperm and oocytes with porcine oviductal epithelial cells on in vitro fertilization.
This study was designed to determine the effect of co-culture with porcine oviductal epithelial cell (POEC) monolayers on in vitro fertilization of pig oocytes. The in vitro penetrability of mature (experiment 1) or immature (experiment 2) oocytes was studied in presence or absence of POEC during IVF with fresh semen. In experiment 3, boar and POEC effects were analyzed but in this case with frozen-thawed spermatozoa. In experiment 4, the spermatozoa were pre-incubated before IVF with or without POEC in order to assess their effect on IVF sperm-related parameters. In experiment 5, the effect of POEC was studied by co-culturing them with oocytes before IVF to determine if monospermy was improved. The results showed that high sperm concentration and POEC increase oocyte penetrability (P<0.01) and decrease monospermy rate (P<0.01), in both mature and immature oocytes (P<0.01) with fresh semen and a 18 h culture time. With frozen semen was detected a boar and POEC effect (P<0.01) on penetration rate. The sperm pre-culture 2 h with POEC also resulted in an increase of sperm penetration in terms of number of sperm per oocyte (P<0.01) and this treatment did not increase monospermy when contact time between gametes was limited to 6 h although monospermy was higher when POEC were present during IVF. Finally, exposure of oocytes to POEC for 4 h before IVF facilitated monospermic penetration to over 70% (P<0.01). In conclusion, the use of POEC in porcine IVF systems provides the possibility of working with low sperm concentrations and the effect of POEC on monospermy depends on sperm concentration, boar and contact time between gametes. Moreover, the exposure of oocytes to POEC before IVF improves the rate of monospermy.
Romar
R
R
Department of Animal Biology (Animal Physiology), Faculty of Veterinary Medicine, University of Murcia, 30071, Murcia, Spain. rromar@um.es
Coy
P
P
Campos
I
I
Gadea
J
J
Matás
C
C
Ruiz
S
S
D000818:Animals; D002478:Cells, Cultured; D018920:Coculture Techniques; D015925:Cryopreservation / Q000662:veterinary; D004847:Epithelial Cells; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D008297:Male; D009865:Oocytes / Q000502:physiology*; D010057:Oviducts / Q000166:cytology / Q000502:physiology*; D012662:Semen Preservation / Q000662:veterinary; D013075:Sperm Capacitation; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000502:physiology*; D013552:Swine / Q000502:physiology*