Liu Jun, Van der Elst Josiane, Van den Broecke Rudy, Dhont Marc
Infertility Centre, Department of Obstetrics and Gynaecology, Ghent University Hospital, De Pintelaan 185, 9000 Ghent, Belgium.
Hum Reprod. 2002 Mar;17(3):605-11. doi: 10.1093/humrep/17.3.605.
Ovarian tissue cryopreservation and transplantation can be used to restore fertility to sterile females. A question that warrants further investigation is whether the follicular content is affected by the freeze-thawing and grafting procedure, and if so, to what extent and by what mechanism.
Intact newborn mouse ovaries were allografted under the kidney capsule or were cryopreserved by slow freezing with dimethylsulphoxide as the cryoprotectant prior to grafting. Estrogenic activity of ovariectomized recipient mice, as revealed by vaginal cytology, resumed after 11 days of transplantation. At 14 days after transplantation, ovarian grafts were recovered and processed histologically for follicle number counting. The follicular content of grafts of fresh ovaries was 58% of that from ovaries of age-matched 14 day old mice. In frozen-thawed ovarian grafts, the follicular content was only 9% lower than that of fresh grafted ovaries. Apoptosis of follicular cells was investigated by DNA nick end labelling. We observed a marked increase in the staining of fragmentation of DNA shortly after transplantation (2-12 h) of fresh newborn mouse ovaries.
The results of the present study indicate that transplantation rather than cryopreservation accounts for the major and early loss of primordial follicles in grafted newborn mouse ovaries.
卵巢组织冷冻保存和移植可用于恢复不育女性的生育能力。一个值得进一步研究的问题是，卵泡含量是否会受到冻融和移植过程的影响，如果是，影响程度如何以及通过何种机制。
完整的新生小鼠卵巢被异体移植到肾包膜下，或者在移植前用二甲亚砜作为冷冻保护剂进行慢速冷冻保存。通过阴道细胞学检查发现，卵巢切除受体小鼠在移植11天后恢复了雌激素活性。移植后14天，回收卵巢移植物并进行组织学处理以计数卵泡数量。新鲜卵巢移植物的卵泡含量是年龄匹配的14日龄小鼠卵巢卵泡含量的58%。在冻融后的卵巢移植物中，卵泡含量仅比新鲜移植卵巢低9%。通过DNA缺口末端标记法研究卵泡细胞的凋亡。我们观察到，新鲜新生小鼠卵巢移植后不久（2 - 12小时），DNA片段化染色显著增加。
本研究结果表明，移植而非冷冻保存是导致移植的新生小鼠卵巢中原始卵泡大量早期丢失的主要原因。
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