The relation with cytoskeletal changes of human ooyte and their age.

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We reported about cytoskeletal changes oocyte and sperm in fertilization. In current study, we tried to take human immature oocytes from patients who underwent gynecological surgery and these oocytes cultured in 37℃, 5%CO2 atmosphere at 24hours to maturate. The culture medium was made from P-1 medium with a little changes given reported by Cekeleniak in 2001. The oocytes were collected from three polycystic ovarian syndrome patients who underwent laparoscopic ovarian dolling. The first case is 25 years old and eight immature oocytes were collected. After culture for maturation at 24 hours, these oocytes changed into six metaphase II (MII) eggs, one germinal vesicle (GV) egg and one degenerate egg. The second case was 26 years old and twelve oocytes were collected. These oocytes changed into two MII eggs, four GV eggs and six degenerate eggs. The third case was 33 years old and four oocytes were collected. These oocytes changed into one MII egg, two GV eggs and one degenerate egg. The rate of successes of maturation culture was 37.5%. These eggs were fixed and observed alpha-beta tubuline and DNA by immunohistochemical method that reported S Simerly and G Schatten. We recognized these spindles were no specific differences from in vitro maturated oocyte in tubuline and chromosome structure to in vivo matured eggs. We tried to observe the real time changes of spindles of culture eggs by poloscope. However, all of eggs that observed by poloscope were degenerated though we could not get any specific changes in this experiment by poloscope. In conclusion, we studied in vitro culture for human immature oocytes into mature eggs, there were no changes in cytoskeletal observation. This culture method will useful for research about human oocyte maturation kinetics. Further, We will study the in vitro culture system to another age of people.