Effect of methyl-beta-cyclodextrin treatment of pig spermatozoa on in vitro fertilization and embryo development in the absence or presence of caffeine.
A series of experiments were carried out to develop a new method to reduce pig polyspermic fertilization and produce more normal embryos, in vitro. Experiment 1 determined the effect of methyl-beta-cyclodextrin (MCD) treatment during cryopreservation on sperm acrosome reaction and sperm fertilization. Compared to the non-MCD-treated control, MCD treatment increased the percentage of acrosome-reacted spermatozoa at thawing and 2h after incubation in fertilization medium (P<0.01). Treatment with MCD also increased (P<0.05) sperm-penetration rate, number of spermatozoa in oocytes, and fertilization efficiency in the caffeine-free fertilization medium. Experiment 2 was designed to examine the effect of withdrawal of caffeine (caffeine-free) from fertilization medium on fertilization parameters and early embryo development. Using MCD-treated spermatozoa, there was no difference in sperm-penetration rate, oocyte cleavage rate, and blastocyst formation rate between the caffeine-free and caffeine-supplemented groups. However, polyspermic fertilization rate was lower, and fertilization efficiency and blastocyst cell number were higher in the caffeine-free group compared to the caffeine-supplemented group (P<0.05). Experiment 3 studied the effect of caffeine and different concentrations of spermatozoa on fertilization parameters. Sperm-penetration rate did not differ between the caffeine-free and the caffeine-supplemented groups at different sperm concentrations. Caffeine and sperm concentration had an effect on the number of spermatozoa in oocytes and on the polyspermic fertilization rate (P<0.002). Caffeine also affected fertilization efficiency (P<0.05). In conclusion, treating spermatozoa with MCD and withdrawing caffeine from fertilization medium may provide a new method to produce a large number of normal embryos, in vitro.
Mao
Jiude
J
Division of Biological Sciences, University of Missouri-Columbia, Columbia, MO 65211, USA.
Wu
Guang-Ming
GM
Prather
Randy S
RS
Smith
Michael F
MF
Cantley
Tom
T
Rieke
August
A
Didion
Brad A
BA
Day
Billy N
BN
D020101:Acrosome Reaction / Q000187:drug effects; D000818:Animals; D014408:Biomarkers, Tumor; D001755:Blastocyst / Q000502:physiology; D002110:Caffeine / Q000008:administration & dosage*; D002970:Cleavage Stage, Ovum / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary; D047108:Embryonic Development; D005306:Fertilization / Q000187:drug effects; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D008297:Male; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary; D013084:Sperm-Ovum Interactions / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*; D013552:Swine; D047392:beta-Cyclodextrins / Q000008:administration & dosage*; D000951:Antigens, Neoplasm
D014408:Biomarkers, Tumor; D002110:Caffeine; D047392:beta-Cyclodextrins; C115012:human epithelial antigen-125; D000951:Antigens, Neoplasm; C108732:methyl-beta-cyclodextrin