| The present study was conducted to investigate the in-vitro maturation (IVM) of human immature oocytes retrieved from small antral follicles of the patients with polycystic ovary syndrome (PCOS) without stimulating, and their competence of fertilization and embryonic development following ICSI. We found that, under the same in-vitro maturation medium (IVMM) systerm as immature oocytes aspirated from stimulated ovaries, those non-stimulated oocytes' in-vitro maturation (71.88%), fertilization (60.87%), cleavage rates (82.14%) and percentage of good-quality embryos on the day 3 after ICSI (6.52%) were significantly lower. The developmental competence of embryos resulted from non-stimulated oocytes after IVM and intracytoplasmic sperm injection (ICSI) was poor, and most of them would be arrested in any stages of embryonic development, especially at the 2- to 8-cell stage. Ultimately, few of them could develop beyond the morula (10.87%) and blastocyst (6.52%) stages. The results show that, there is no significant difference in humam immature oocytes retrieved from non-stimulated and stimulated ovaries with regard to nuclear maturation rate, and when cultured in the present in-vitro maturation medium (IVMM) system, non-stimulated oocytes have ability to resume the meiotic maturation, but subsequently arrest at metaphase- II (M- II) stage. And, their capacity of fertilization and embryonic development were impaired, which should mainly be due to suboptimal cytoplasmic maturation. In addition, the IVMM systerm used in the current study couldn't entirely mimic the dynamic in-vivo physiological environment of oocyte growth and devolpment in the follicle in non-stimulated ovaries. According to the maturity of oocytes and their follicular status at the time of aspirating, weshould design a multi-IVMM system, which could contribute to optimal cytoplasm maturation.To determine the predictive value of oocyte morphologic characteristic on the ability of fertilization and embryo development, all oocytes from ICSI cycles were grouped based on their morphology, including the features of first polar body, perivitelline space and cytoplasmic inclusions, and the fertilization, cleavage rates and percentages of good-quality embryos on the day of embryo transfer were compared among groups. Significant difference among groups according to the first polar body and perivitelline space was observed with regard to fertilization rate(P=0.044). The percentage of good-quality embryos from oocytes without cytoplasmic inclusions was significantly higher than those from oocytes with cytoplasmic inclusions (P=0.011). Present data indicates that, oocyte morphology, including the shape of the first polar body and the size of perivitelline space, reflects the maturity of the oocytes and may have a predictive value on the ability of fertilization following ICSI. Oocytes with the optimal maturity should have a normal perivitelline space, an intact ovoid or round first polar body with a rough surface, and may have more potential for fertilization. Cytoplasmic inclusions are correlated with the degree of cytoplasmic maturation, and are likely to be predictive markers of the competence of embryonic development following fertilization.In the current study, methods of isolating human inner cell masses (ICM), condition for human embryonic stem cells (ESC) culture and passage technique were explored. The poor-quality human embryos, donated by consenting couples undergoing in vitro fertilization (IVF) or ICSI treatment, were used for establishment of human embryonic stem cell (ESC) lines. The results showed that embryo quality is positively correlated with the formation of primary human ICM clones. The ability of blastocyst formation of these poor-quality embryos is compromised,which limits the success in establishing human ESC lines. We isolated the pure ICM from the trophectoderm with two methods, and found that the efficiency of isolating would be higher by immunosurgery than by culturing the hatched blastocysts directly. In our study, we establis...
|
PDF Full Text Request