Studies On Zygote Ivc After ICSI Using Abnormal Or Weak Sperm

Recently, the harms of environmental estrogen to male anmimals and the ICSI technology's effect of causing abnormal to embryo development are still disputed. This experiment was designed based on these disputations.Took advantages of experimental animal carrier, sprinkled β -cypermethrin,which a kind of pyrethrum ester pesticide that had been proved to have the environmental estrogen effect ,on the forage grass and feed them to male rabbits in order to induce abnormality artificially.Using artificial vagina collect the semen, after being treated properly, injected the abnomal sperm or inactive sperm into the cytoplast of mature oocyte by microoperation. Selected the zygotes which had vented the polar body â…¡ or had two primary nucleus to proceed further incubation, research the best conditions to obtain the high rate of fertilization,cleavage and development. This research could be helpful to improve the ICSI zygotes'development, and the results could suggest whether the male infertile patients could have offsprings using the ICSI technology, so could provide basic information and technological reference for the treatment of male infertility, animal breeding and breeding protection of rarity animals.This experiment was mainly done in three aspects: firstly, collected four-month healthy male rabbits with symmetrical weight(1.8±0.10kg),expelled parasite, isolated and observed two weeks, then divided them into four groups randomly, sprinkled the β-CP on the forgae grassa and feed them to the rabbits directly.let each group's pesticide quantity as 0mg.kg^-1d^-1,1.5 mg.kg^-1.d^-1,3.0 mg.kg^-1.d^-1 and 4.5 mg.kg^-1.d^-1 56 days later, collected the semen by artificial vagina, proceed routine analysis with the semen to assay the pesticide's effect of causing abnormality.Secondly, six-month healthy young female rabbits without pregnancy whose weight were above 2.0kg-3.5kg, dealed them with PMSG+HCG for superovulation and mated them with normal male rabbits in the same cage. After 19-20h, obtain the embryo by surgery, cleaning them, and then puting them in three different culture liquid as TCM199+ 10%NBS,RPMI1640+10%NBS,RPMI1640+10%FBS respectively. Under the same condition (38.5 ℃ ,5% CO₂,saturant humidity) cultured the pronulear embryos continually, observing the state of in vitro development,at last confirming the dominant culture system for rabbits' zygotes. Thirdly, the first-group rabbits'semen were collected by artificial vagina.Diluted the semen and washing them by centrifugal effect,then storing them(some would be dealed with DM liquid to make sperm capacitation,some would...