Effects Of Antioxidants On The Mouse Parthenogenetic Embryo Development In Vitro

Objective The purpose of this study is to explore the effect of different concentrations of glutathione(GSH),vitamin E(VE),vitamin C(VC)on the development of mouse parthenogenetic embryos in vitro.And determine the optimal concentration of three antioxidants to promote the development of mouse parthenogenetic embryos in vitro,and provide experimental basis for improving embryo culture system and development potential of parthenogenetic embryo in vitro.Methods Selecting the CD-1? female mice as the research object,according to the different antioxidants were divided into three experimental groups,each set of 4concentration points,10 female mice were randomly assigned to the each concentrations point.The GSH group set up 0mmol/L(blank control group),0.5mmol/L,1mmol/L,1.5mmol/L four concentration point to test;Vitamin E group set up the 0μmol/L(blank control group),50umol/L,100umol/L,150umol/L four concentration point to test;Vitamin C group set up the 0μmol/L(blank control group),50umol/L,100umol/L,200umol/L four concentration point to test.Collect the oocytes after ovulation induction in mice and parthenogenetic activate the oocytes in vitro.The activated oocytes were randomly assigned to culture medium containing glutathione,vitamin E and vitamin C,and cultured in incubator at 37℃,5% CO2 and saturated humidity,and the cleavage rate,blastocyst formation rate and blastocyst rate were observed and counted,The effects of antioxidants on the microstructure of parthenogenetic embryos were observed by transmission electron microscopy at the cleavage and blastocyst stages.Results 1 In the glutathione(GSH)group,the concentration of mouse parthenogenetic cleavage rate show no significant difference(P>0.05),blastocyst formation rate and blastocyst hatching rate was statistically significant differences(P<0.05);When concentration was 1mmol/L,developmental potential in every embryonic stages was the best,the cleavage rate,blastocyst rate and blastocyst hatching rate were the highest.Under the transmission electron microscope observation,we could find glutathione group with good structure and few mitochondrial vacuoles and apoptotic cells compared with the control group.2 In the Vitamin E(VE)group,the each concentration of mouse parthenogenetic cleavage rate,blastocyst formation rate and blastocyst hatching rate was statistically significant differences(P<0.05);When concentration of VE was 100μmol/L,the developmental potential in every embryonic stages was the best,the cleavage rate,blastocyst rate and blastocyst hatching rate were the highest.Under the transmission electron microscope observation,we could find VE group with good cell microstructure and few mitochondrial vacuoles and apoptotic cells compared with the control group.3 In the Vitamin C(VC)group,the each concentration of mouse parthenogenetic cleavage rate,blastocyst formation rate and blastocyst hatching rate was statistically significant differences(P<0.05);When concentration of VC was 200μmol/L,the cleavage rate,blastocyst rate and blastocyst hatching rate were the highest.When concentration of VC was 300μmol/L,the cleavage rate,blastocyst rate and blastocyst hatching rate were the lowest.Under the transmission electron microscope observation,we could find200μmol/LVE group with good cell microstructure compared with the control group,and300μmol/LVE group with more mitochondrial vacuoles and apoptotic cells compared with the control group.4 In the 1.0mmol/L glutathione,100μmol/L Vitamin E and 200μmol/L Vitamin C experimental group,200μmol/L VC group obtained cleavage rate,blastocyst formation rate and blastocyst hatching rate were the highest,compared with other different concentration of antioxidants were statistically significance(P<0.05).Conclusions 1 The addition of a certain concentration of antioxidants in embryo culture medium could promote the development of mouse parthenogenetic embryos in vitro.1mmol/L GSH,100 μmol/L VE and 200μmol/L VC were the suitable concentration to improve the developmental potential.2 300μmol/L VC significantly reduced the developmental potential of mouse parthenogenetic embryos,suggesting that high concentrations of VC may have adverse effects on parthenogenetic embryos.3 Three different concentrations of antioxidants,200 μmol/L VC was able to improve the development of mouse parthenogenetic embryos in vitro obviously,and obtained higher cleavage rate,blastocyst formation rate and blastocyst hatching rate.