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

我们报道了受精过程中卵母细胞和精子的细胞骨架变化。在本研究中，我们尝试取妇科手术患者的人未成熟卵母细胞，将这些卵母细胞在37℃，5%CO2的气氛中培养24小时使其成熟。培养基由P-1培养基制成，并在2001年Cekeleniak的报道中做了一些改动。卵母细胞采集自3例多囊卵巢综合征患者行腹腔镜卵巢穿刺。第一例患者25岁，收集了8个未成熟卵母细胞。24小时培养成熟后，这些卵母细胞转变为6个中期II （MII）卵，1个生发囊泡（GV）卵和1个退化卵。第二例患者26岁，采集了12个卵母细胞。这些卵母细胞变成了2个MII卵、4个GV卵和6个退化卵。第三例患者33岁，采集到4个卵母细胞。这些卵母细胞变成了一个MII卵、两个GV卵和一个退化卵。成熟培养成功率为37.5%。用S Simerly和G Schatten报道的免疫组织化学方法对这些卵进行固定，观察α - β小管蛋白和DNA。我们认识到这些纺锤体在体外成熟卵母细胞的微管和染色体结构上与体内成熟卵没有特异性差异。我们尝试用马孔镜观察培养卵纺锤体的实时变化。然而，poloscope观察到的所有卵子都发生了退化，尽管我们在这次实验中没有看到任何具体的变化。综上所述，我们所研究的体外培养将人未成熟卵母细胞转化为成熟卵，观察到细胞骨架无变化。这种培养方法对人类卵母细胞成熟动力学的研究具有重要意义。进一步，我们将研究体外培养系统到另一个年龄的人。

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.