The Research On The In-vitro Maturation And Vitrification Of Oocytes From Human Luteal Phase

ObjectiveTo investigate in vitro maturation, vitrification-thawing and ICSI of immature oocytes from human luteal phase as to preserve the fertility of some female cancer patients.MethodsImmature oocytes (n=256) were aspirated from women undergoing gynecologic operations in Gynecology and obstetrics department of Qilu Hospital of Shandong University from May2010to May2011. They were divided by menstrual cycle, into controlled group and virtrification group. The oocytes of controlled group were matured in vitro and then inseminated by intra cytoplasmic sperm injection. The oocytes of virtrification group were matured in vitro to MⅡ, following virtrification, then thawed and inseminated by intra cytoplasmic sperm injection. The oocytes were cultured with their attached compact surrounding cumulus cells in maturation medium which contained Tissue Culture Medium199(M199, Sigma, USA) which supplemented with0.075IU/ml FSH(rFSH, Gonal-F, Serono),2IU/ml HCG,10%(v/v) HFF,10%(v/v) SSS (Irvine Scientific, USA) and0.2ug/ml17-oestradiol (Sigma). Maturation rate, survival rate, fertilization rate, cleavage rate and rate of high quality embryos were compared among the groups.The times of maturation in vitro were also recorded.Results1. There was no significant difference in MⅡ stage rate between oocytes retrieved at luteal phase and follicular phase. The survival rate,68.0%VS48.1%, was higher for the oocytes retrieved at follicular phase than at luteal phase (P<0.05). The fertilization rate, cleavage rate and rate of high quality embryos showed no significant difference between the two groups.2. Cryopreservation showed significant effect on the fertilization rate and cleavage rate in both of the two groups.3. The times of maturation in vitro is from24h to72h,120h at the longest. The oocytes cultured24h、48h、72h were accounted for25.4%(65/256),52.7%(135/256),19.5%(50/256) respectively.Conclusion1. Immature oocytes retrieved at luteal phase can be matured in vitro and developed to high quality embryos.2. Vitrification declines the fertilization rate and cleavage rate of oocytes.3. In vitro maturation, vitrification-thawing and intra cytoplasmic sperm injection of immature oocytes is a convenient way for female cancer patients to preserve their fertility in the clinical work.