Wang Chia-Woei, Lai Ying-Ming, Chan Ping-Ru, Horng Shang-Gwo, Chang Chia-Lin, Chen Chun-Kai, Wu Hsien-Ming, Huang Hong-Yuan, Wang Hsin-Shih, Soong Yung-Kuei
Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, Lin-Kou Medical Center, Kwei-Shan, Tao-Yuan, Taiwan.
J Assist Reprod Genet. 2002 Oct;19(10):493-9. doi: 10.1023/a:1020366620890.
Ovarian tissue banking may be the best strategy to preserve female fertility. But optimal method to obtain viable mature oocytes remains challenging. In order to bypass the long in vitro oocyte growth period, we developed this study to test whether reconstruction of thawed primordial oocytes with enucleated preovulatory germinal vesicle (GV) oocytes could induce dictyate nuclei to undergo chromosomal condensation and meiotic maturation.
Isolated primordial oocytes from thawed mouse ovarian tissue were reconstructed with enucleated GV oocytes. After electrofusion and in vitro maturation, the reconstituted oocytes were assessed for first polar body extrusion, cytoskeleton configuration, and chromosome abnormalities.
Primordial oocytes from thawed ovarian tissue showed a high survival rate. Following transfer and electrofusion, they could be fused with enucleated GV oocytes (35.6%, 36/101) and extruded a first polar body (52.8%,19/36). These mature oocytes showed a normal spindle configuration and chromosome number.
We successfully established a mouse cell model to prove that omitting the whole growth and maturation period by transfer of primordial oocytes to developmentally older enucleated oocytes would bypass the long growth period required to the preovulatory stage. Polar body extrusion could also ensue after in vitro growth. This study provided an alternative approach for future investigations in oocyte maturation.
卵巢组织冻存可能是保存女性生育力的最佳策略。但获取有活力的成熟卵母细胞的最佳方法仍具有挑战性。为了绕过卵母细胞漫长的体外生长阶段，我们开展了本研究，以测试用去核的排卵前生发泡（GV）卵母细胞重建解冻后的原始卵母细胞是否能诱导双线期细胞核发生染色体浓缩和减数分裂成熟。
用去核的GV卵母细胞重建从解冻的小鼠卵巢组织中分离出的原始卵母细胞。经电融合和体外成熟后，对重构卵母细胞进行第一极体排出、细胞骨架结构和染色体异常情况的评估。
解冻卵巢组织中的原始卵母细胞显示出较高的存活率。在移植和电融合后，它们能够与去核的GV卵母细胞融合（35.6%，36/101）并排出第一极体（52.8%，19/36）。这些成熟卵母细胞显示出正常的纺锤体结构和染色体数目。
我们成功建立了一个小鼠细胞模型，证明将原始卵母细胞移植到发育更成熟的去核卵母细胞中，省略整个生长和成熟阶段，可以绕过排卵前阶段所需的漫长生长时期。体外培养后也可出现极体排出。本研究为未来卵母细胞成熟的研究提供了一种替代方法。
