Effect of methyl-beta-cyclicdextrin(MBCD)on porcine in vitro fertilization and early embryo development

Lü Ming*,HE Wen-teng*,JIAO Ming-xia,HUAN Yan-jun, SHI Yong-qian,KONG Qing-ran,LIU Zhong-hua**(College of Life Science,Northeast Agricultural University,Harbin 150030,China)

methyl-beta-cyclic dextrin;in vitro fertilization;sperm capacitation;pig

In order to optimize the porcine IVF technology system,this experiment was designed to explore the influence of methyl-beta-cyclic dextrin(MBCD)on porcine in vitro fertilization and early embryo development.Different concentrations(0,0.5,1,2,5,10,15,20 μmol/mL) of MBCD was added to modified Tris-buffered medium(mTBM) at 0 h and 4 h of in vitro fertilization then the resulted embryos were cultured in PZMμ3.Embryo in vitro development rate and polyspermy proportion was examined,and sperm capacitation state was assessed by chlortetracycline(CTC) staining.Results:(1)In the group of adding 5 μmol/mL MBCD to mTBM at IVF 0 h the cleavage rate,blastocyst rate,cell number of blastocyst was significantly(P<0.05) higher than that in the control group and the other experimental group except 10 μmol/mL group;(2)In the group of 5 and 10 μmol/mL MBCD to mTBM at IVF 0 h the monospermy rate was significantly(P<0.05) higher than that in the control group and the other experimental groups,and the polyspermy rate in group of 5 and 10 μmol/mL MBCD was significantly(P<0.05) lower than that in the control group and the other experimental groups;(3)In the group of 5 μmol/mL MBCD to mTBM at IVF 0 h,during 0-1 h of incubation,the number of F type sperm rapidly reduced(78.56-19.43);while the number of B type sperm increased rapidly(10.79-69.86).During 1-4 h of the incubation,the number of F type and B type sperm had not much change(B: 69.86-78.78,F: 19.43-9.11).In conclusion,the addition of 5 μmol/mL MBCD at 0 h of IVF can significantly improve capacitation sperm percentage,reduce polyspermy fertilization,and thereby enhance the early embryo development competence.