冻存复苏人卵巢皮质在裸鼠体内的发育及卵细胞的提取(英文)

Oocyte retrieval and in vitro maturation from frozen thawed human ovarian cortex transplanted in immune suppressed mice

目的探求冻存复苏人卵巢皮质在裸鼠体内的发育及卵细胞的提取。方法取手术切除的5例患者的卵巢皮质,放入1.5mol/L的二甲基亚砜+10%血清中,应用可控程序冻冰仪逐渐降温,移入液氮中冻存。3个月后,将融解的卵巢皮质移植入90只裸鼠皮下,注射hFSH12周,刺激卵泡生长。注射hCG36h后,用18号针头穿刺提取卵细胞。结果卵巢直径为3～9mm,穿刺获19个卵细胞,放入TC-199培养基,加20%胎牛血清,25mmol/L丙酮酸,75mIU/mLFSH和LH培养,获15个分裂中期的次级卵母细胞。结论人卵巢组织冻融后可异体移植生长,且其卵细胞可在体外发育成熟。

[Objective] Storage of ovarian tissue may restore long-term fertility in patients undergoing treatment that irreversibly damages the ooeyte population. The potential of transplantation of frozen-thawed human ovarian tissue for ooeyte retrieval and maturation was investigated. [Methods] Ovarian tissues from 5 patients were surgieally removed, disseeted, frozen in 1.5 mol/L DMSO＋10% serum using a programmable freezer and transferred to liquid nitrogen. Three months later, 89 nude miee were transplanted with thawed ovarian eortex fragments, stimulated using hFSH injections for 12 weeks and ooeyte retrieval were performed 36 hours post-hCG using 18-gauge needles. [Results] 19 oocytes were obtained from 3 to 9 mm follicles and 15 of them were matured in-vitro （IVM）, in TC- 199 medium with 20% fetal bovine serum, 25 mmol/L of pyruvie acid, and 75 mlU/mL of FSH and LH, to the metaphase stage. [Conclusion] We eonelude that human ovarian follicles survive freeze-thaw and long-term xenograft transplantation. Ooeyte retrieval and maturation from transplanted frozen human ovarian cortex into mice is feasible.