Rearches On Round Spermatid Injection (ROSI) And Relating Problems In Porcine

The appearance of Round spermatid injection (ROSI) provides a new way to treat the sterile human male . As an new animal embryo biotechnology, ROSI can raise the animal gamete utilizing efficiency, gain certain sex individuals, the solution productivity difference animal fertilization question, as well as transfers the genetic engineering in the achievement the auxiliary technical aspect all to have the important application. The ROSI technology studies at present is not thorough, the clinical practice success ratio is extremely low, but also is in tries to find out the stage, in the animal, merely in the mouse, the rabbit, the monkey and so on has the report in particular, not yet sees the report in porine.The porine is the polyembryony animal, many obtains the ovocyte from the porine ovary although but the maturity quite to be low; Because the ROSI technology is does not have to grow the mature sole round sperm cell to pour into directly completely in the ovicell nature, jumped over the spermatozoon in to pass through the physiology and the biochemistry, if the ovicell nature mature or the activation degree were insufficient, added the round immature sperm cell maturity quite inferior reason, very possibly Causes the ROSI micro fertilization defeat. This experiment is for the purpose of unifying the porine ovocyte in vitro mature raise and the ROSI correlation operation, establishes in the ovicell nature the round sperm cell microinjection fertilization technology procedure, the exploration porine round sperm cell micro fertilization possibility, the research affects this kind of special cell microinjection fertilization effect the factor, for further studies the porine ROSI technology to provide the basic data.This experiment first studied in the mature nutrient medium to increase the hormone to the porine ovocyte in vitro mature process in the chromosome configuration and the mature rate influence, by explored the porine ovocyte in vitro mature rule, enhanced in vitro raise maturity, was the ROSI technology system preparation high quality ovocyte; Then has carried on the experiment in view of the porine round sperm cell in vitro separation, through to the ROSI operation and ICSI (Intracytoplasmic sperm injection, the comparison which ICSI) operated studies has affected the ROSI operation the factor. Findings as follows:Experiments 1: Hormone to ovocyte IVM mature ratio influence: Dyes with H33342 observes COCs in three kind of different culture environment (FSH → without hormone,FSH→ LH, FSH + LH → without hormone. "→ " The expression trades the medium in cultured 22th ~ 24 h). After culture 48h, FSH → without hormone group GV rate proportion remarkably to be higher than other two groups (22.73% vs 9.86%, 4.95%, P<0.05); FSH + LH → without hormone group the MII...