Studying The Incubator In Different Concentrations Of Ethanol And Chloroform In Mice Of The Effect Of In Vitro Fertilization And Embryonic Development

Objectives The purpose of this study is to explore the different concentrations of ethanol and chloroform in incubator in mice in vitro fertilization and embryo development.Through access to observe and record the different concentrations of ethanol and chloroform between groups of the fertilized egg number, number of cleavage, blastocyst and blastocyst hatch results, calculate the rate of fertilization, cleavage, blastocyst formation and blastocyst hatched for human assisted reproductive technology technical standardization and to provide experimental basis to improve success rate of human IVF-ET.Methods 8 to 12 weeks of age female and male mice of kunming mice were chosen as the research object. After stimulating ovulation with Female mice, the eggs of female mice in vitro fertilization and embryo culture were removed. We bubbled into ethanol and chloroform, two gases experiment in the incubator respectively, and set up blank control group, and four groups of experiments with 5mg/m~3, 10mg/m~3, 20mg/m~3,and 40 mg/m~3 of the concentration of ethanol gas respectively. In addition, We set up blank control group and four groups of experiments that they are 10mg/m~3, 20mg/m~3, 40mg/m~3,and 80mg/m~3 of the concentration of chloroform gas group. The above experiment in each group randomly assigned to eight female and eight male mice. Access to observe and record the different concentrations of ethanol and chloroform gases between groups of the fertilized egg number, number of cleavage, blastocyst and blastocyst hatch results. Statistical analysis for different concentrations of ethanol and chloroform in mice in vitro fertilization and embryo development impact was conducted. In addition, random sampling was done for each group of experimental oocytes, observing its spindle morphology by confocal laser technology.Results 1 Compared respectively ethanol gas concentration of 5,10, 20 and 40mg/m~3 with the blank control group, the mouse oocyte in vitro fertilization rate is respectively(91.2%,89.9%, 89.0%, 86.8%, 73.2%) and cleavage rate is respectively(85.8%, 85.9%, 84.8%,84.1%, 83.5%), the blastocyst formation rate is respectively(85.0%, 82.8%, 81.3%, 80.2%,59.4%) and blastocyst hatched rate respectively(79.6%, 78.2%, 76.0%, 74.2%, 40.4%);Cleavage rate after comparing different concentrations of no statistical difference(P >0.05); And fertilization rate, blastocyst formation rate and blastocyst hatched rate were statistically significant, and is characterized by a linear trend, namely with concentration increasing, all into decline(linear correlation(P < 0.05). Ethanol gas concentration is 40mg/m~3 oocytes appear degeneration, breaking of the spindle. 2 Compared respectively chloroform gas concentration of 10, 20,40 and 80mg/m~3 with the blank control, the concentration of chloroform of 0,10,20,40 and 80 mg/m~3 of mouse oocyte fertilization rate is respectively(89.0%, 87.3%, 86.4%, 85.6%, 71.6%) and cleavage rate respectively(83.6%, 83.3%, 82.9%, 81.8%, 64.7%), the blastocyst formation rate is respectively(82.0%, 80.9%, 79.3%, 60.7%, 53.3%) and blastocyst hatched rate respectively(67.0%,65.6%, 64.1%, 35.3%, 25.0%); Comparing different concentrations after fertilization rate,cleavage rate, blastocyst formation rate and blastocyst hatched rate were statistically significant(P < 0.05), and is characterized by a linear trend, namely with concentration increasing, into a downward trend(linear correlation(P < 0.05). Chloroform gas concentration of 80 mg/m~3 degeneration of the spindle, the fracture occurs.Conclusions 1 Along with the increasing concentration of the ethanol gas in the incubator, the mouse oocyte fertilization rate in vitro fertilization and blastocyst formation rate and blastocyst hatched rate are on the decline. 2 Along with the increasing concentration of chloroform in the incubator of the gas, the mouse oocyte fertilization in vitro fertilization rate, cleavage rate, blastocyst formation rate and blastocyst hatched rate are all on the decline. 3 Controlling the concentration of ethanol and chloroform in the incubator under 20 mg/m~3,the influence of in vitro fertilization and embryo development is not obvious.