The Research Of Technique Of Mouse Embryos Culture In Vitro

Objective: To evaluate the effect of different culture medium on mouse embryos culture, then search the best culture system of embryos in vitro, in order to provide a theory that can improve human embryos culture, and promote the outcomes of in vitro fertilization-embryo transfer(IVF-ET).Methods: As a supplement, human follicular fluid (HFF), serum and decidua conditioned medium were added to Earle respectively. 1678 mouse ovums were fertilized in different culture medium, and then 2-cell mouse embryos were cultured in these medium. And the following changes of the above system were closely observed under microscope. The fertilization rate, cleavage rate and speed, good embryo rate, morula rate and blastocyst rate were analyzed.Results:一 1, Among the embryos cultured in decidua conditioned medium, there were 72.9%, 66.0%, 61.6% and 32.1% developed to 8-cell stage, morula stage, blastocyst stage and hatching, there are significant difference compared with control(43.6%, 30.9%, 19.1%, 5.5%.P<0.01). 2, HFF significantly increased frequency of development from the 2-cell stage to the 8-cell stage(71.3%), morula stage(58.8%), blastocyst stage(50.0%) and hatching(25.7%) compared with control(P<0.01). 3, there are no different about the number of each stage of embryos between serum group(53.4%, 40.5%, 29.3%, 7.8%) and control(P>0.05).二 There are no difference among different medium about the fertilization rate. However, about cleavage rate and good embryo rate, significant difference were found between HFF and control or between decidua conditioned medium and control, but there are no difference between serum and contol. Embryos cultured in HFF group and decidua conditioned medium group cleaved significantly faster than control and showed no or lessfragmentation than those in the control.三 HFF group and decidua conditioned medium group compared with control, significant different were found in embryo degeneration, on the contrary, there are no difference in embryo degeneration between serum and control.四 When HFF and decidua conditioned medium was added to serum group respectively, the development of embryo is significantly increased. Compared with semm group, there have both significant difference in each stage between these group and serum group (P<0.01).Conclusion: These results suggested that decidual cell can release some substances that can support early embryonic development and yield a reasonable number of embryos with good quality up to the blastocyst stage. And human follicular fluid is effective in increasing the developmental potential of mouse embryos in vitro.