Telfer Evelyn E, McLaughlin Marie, Ding Christina, Thong K Joo
Institute of Cell Biology, The Darwin Building, University of Edinburgh, The King's Buildings, Mayfield Road, Edinburgh EH9 3JR, UK.
Hum Reprod. 2008 May;23(5):1151-8. doi: 10.1093/humrep/den070. Epub 2008 Mar 6.
The objective of this study was to determine whether follicles grown within human ovarian cortical strip culture for 6 days in serum-free medium could be isolated at the secondary stage of pre-antral development and grown in vitro to the late pre-antral/early antral stage during a 4 day culture period.
Ovarian cortical biopsies were obtained from six women aged 26-40 years, with informed consent, during elective Caesarean section. Small tissue slices of ovarian cortex, with underlying stromal tissue removed, were cultured in serum-free medium for 6 days and at the end of this period pre-antral (secondary) follicles were dissected from the strips. Seventy-four intact pre-antral follicles ranging in size (66-132 microm) (mean size 100 microm +/- 3.4) were selected for further culture. Follicles were placed individually within V-shaped microwell culture plates in serum-free medium in the presence (n = 38) or absence (n = 36) of 100 ng/ml of human recombinant activin A.
Pre-antral follicles grown for 4 days in the presence of activin A grew to a larger size (mean diameter 143 microm +/- 7.4) than those grown in control medium (mean diameter 111 microm +/- 8) (P < 0.005). Ninety percent of follicles cultured in the presence of activin A increased in size during the first 2 days of culture compared with only 36% of follicles in control medium (P > 0.005). Of the follicles surviving the entire culture period, 30% of those cultured in the presence of activin A showed normal morphology with intact oocytes and antral formation. None of the follicles grown in control medium developed antral cavities and >90% of those follicles collected at the end of the culture period showed signs of oocyte degeneration.
The results reported here demonstrate that under certain conditions, it is possible to achieve accelerated oocyte/follicle development from human primordial/primary follicles. This provides the first encouraging step towards achieving full in vitro growth of human oocytes.
本研究的目的是确定在无血清培养基中培养6天的人卵巢皮质条内生长的卵泡是否能在窦前发育的次级阶段分离出来，并在4天的培养期内体外生长至窦前晚期/早期窦状卵泡阶段。
在择期剖宫产时，经六名年龄在26 - 40岁的女性知情同意后获取卵巢皮质活检组织。去除卵巢皮质下方的基质组织后，将小组织切片在无血清培养基中培养6天，在此培养期末，从皮质条中解剖出窦前（次级）卵泡。选择74个完整的窦前卵泡，大小在66 - 132微米之间（平均大小100微米±3.4）用于进一步培养。将卵泡分别置于V形微孔培养板中，在含有（n = 38）或不含有（n = 36）100 ng/ml人重组激活素A的无血清培养基中培养。
在激活素A存在的情况下培养4天的窦前卵泡比在对照培养基中培养的卵泡长得更大（平均直径143微米±7.4）（平均直径111微米±8）（P < 0.005）。在激活素A存在的情况下培养的卵泡，90%在培养的前两天体积增大，而对照培养基中只有36%的卵泡体积增大（P > 0.005）。在整个培养期存活的卵泡中，在激活素A存在的情况下培养的卵泡中有30%显示出正常形态，卵母细胞完整且有窦腔形成。对照培养基中培养的卵泡均未形成窦腔，培养期末收集的卵泡中>90%显示出卵母细胞退化的迹象。
此处报道的结果表明，在某些条件下，有可能使人原始/初级卵泡的卵母细胞/卵泡发育加速。这为实现人卵母细胞的完全体外生长迈出了令人鼓舞的第一步。
