Age impact on chromosome and spindle disorganizations in MI and MII oocytes

Oocyte aneuploidies are increased with advancing age in human and these aneuploidies are associated with maternal ageing and result from non-disjunctions and meiotic errors. The objective of this work was to use mouse as a model to study the effect of aging on chromosome and spindle disorganizations in MI and MII CBA/Caj mouse oocytes. Mice were divided to young, [3 months] and aged, [8 months] groups and Swiss Webster mice were used as a double control. The animals were superovulated and the oocytes were retrieved in FHM medium 16h after HCG injection. Immature [MI] and mature [MII] ovulated oocytes were recovered from ampulla and fixed, washed in block-wash solution. Spindle microtubules and chromosomes were visualized using immunofluorescent microscopy with anti beta tubulin monoclonal antibodies and staining with propidium iodide, respectively. The chromosome disorganization rate in MI young oocytes was lower than MII oocytes. The correlation between chromosome and spindle disorganizations was significantly higher in MI and MII oocytes [P<0.05] compared to control. The percentage of the MII oocytes was significantly higher [P<0.01] compared to MI oocytes in the young group but this was not the case in the aged group. This may indicate that the oocyte maturation mechanism was affected adversely by ageing process. In conclusion, the positive correlation between chromosome and spindle disorganizations in MI and MII oocytes may be an indicator for embryo aneuploidy. Ageing induced an increase in the production of ovulated MI versus MII oocytes. CBA/Caj mouse may be used as animal model for study ageing effect on reproductive system

Pregnancy outcome in the luteal defective infertile women following ICSI, embryo transfer and luteal support therapy

Defects in the formation and function of the corpus luteum result in a reduction of progesterone secretion and inadequate secretory transformation of the endometerium. Luteal defective cycles are recognized in 50% of unovulatory patients and 60% of women with recurrent abortions. The goal of the present study was to study the outcome of ICSI in infertile women with mild and marked luteal defective cycles following embryo transfer. Patients and Methods: Women with mild and marked luteal defective cycles were undergone ovulation induction, oocyte retrieval, ICSI and embryo transfer program. Follicular fluid progesterone, estradiol, testosterone were measured. The pregnancy rates in both luteal defective cycles [LDC] were recorded. The concentration of estradiol and progesterone were significantly higher in the mild LDC compared to marked LDC. The concentration of prolactine and testosterone were significantly higher in the marked LDC than mild LDC. The concentrations of FSH and LH were not significantly different between both groups of LDC. The ICSI rate was 74.4% in the mild and 70.7% in the marked LDC. The embryo transfer rate was significantly higher in the mild LDC compared to the marked LDC group [85.2% versus 50% respectively, P<0.001]. The pregnancy rate per embryo transfer was 30.3% in the marked LDC compared to 38% in the mild LDC group. The patient cancellation rates were 13% and 4% in the marked and mild LDC groups. The application of ICSI and luteal support therapy improves embryo implantation and pregnancy rates in infertile women with mild and marked luteal defective cycles and in men with severe teratozoospermia and poor sperm penetration score

In vitro fertilization of immature oocytes by testicular sperm: animal model for azoospermic infertile patients

Men with azoospermia require testicular and epididymal sperm aspiration for intracytoplasmic sperm injection. Women over 37 years of age demonstrate an increase number of immature oocytes after induction of ovulation. The development of a technique for in vitro maturation [IVM] and fertilization [IVF] of the oocytes using testicular, epididymal and vassal sperm [TS, ES, VS, respectively] will be of therapeutic value for the treatment of azoospermic patients. The goal of the study was to develop an animal model for the treatment of infertile couples with obstructive azoospermia and immature oocytes. Canine ovaries and testes were collected from a local animal clinic. Immature oocytes were retrieved from the ovaries and cultured in modified tissue culture medium [MTCM]. The mature oocytes were in vitro fertilized by TS, ES, and VS. The normality of the fertilized oocytes was studied. Sperm motility index was significantly higher [P<0.01] in sperm retrieved from the vas deference compared to ES and TS. The concentration of the sperm was significantly higher [P<0.05] in the testes compared to epididymis and vas deference while VS and ES normal morphology were significantly [p<0.01] better than TS. Viable oocyte percentage was significantly higher [P<0.05] in 72 hours versus 96 hours culture durations. Significant higher IVF was reported in VS compared to other groups [P < 0.05] and the SPI was significantly higher after 96 versus 72 hours duration [P<0.05]. IVM of the oocytes in MTCM for 72 and 96 h resulted in a significant increase in IVM rate compared to 48,120, and 144 h durations. In vitro viable mature oocytes showed higher IVF rates when fertilized with VS compared to ES and TS. The results showed that canine oocytes are good model for azoospermic patients