卵巢组织玻璃化和移植面临的主要挑战包括通过低温保存和缺血再灌注损伤引起的卵泡凋亡，以及移植后的卵泡丢失。本研究旨在调查促卵泡生长激素（FSH）和1-磷酸鞘氨醇（S1P）对冷冻保存和移植后卵巢的保护作用及其潜在机制。将21天龄小鼠的卵巢分别用0.3 IU/mL FSH、2 µM S1P和0.3 IU/mL FSH + 2 µM S1P的玻璃化液保存，并进行卵泡计数和凋亡相关指标的检测。结果表明，在玻璃化过程中进行FSH和S1P联合处理能够通过抑制细胞凋亡来保持原始卵泡库，抑制卵泡萎缩。解冻的卵巢移植到切除卵巢的6-8周龄小鼠的肾上腺下囊，移植后24小时或7天取出。结果表明，FSH和S1P联合处理可以在移植后的24小时内抑制卵巢凋亡和自噬，通过上调Cx37和Cx43表达促进卵泡存活，在移植卵巢中通过促进VEGF表达增强血管生成，并且在移植后7天提高E2水平，恢复卵巢内分泌功能。通过CoCl2处理在人类颗粒样肿瘤细胞系（KGN）建立了低氧缺血细胞模型，通过无血清培养体系进行缺血实验。结果证实，FSH和S1P联合处理可以减少缺血低氧引起的卵巢颗粒细胞凋亡，通过上调AKT/mTOR信号通路来抑制颗粒细胞自噬。总之，FSH和S1P联合给药能够在卵巢玻璃化过程中维持卵巢的存活，增加移植后的卵泡存活和血管生成。© 2023. BioMed Central Ltd., part of Springer Nature.

Some major challenges of ovarian tissue vitrification and transplantation include follicle apoptosis induced by cryopreservation and ischemia-reperfusion injury, as well as ovarian follicle loss during post-transplantation. This research aimed to investigate the protective effects and underlying mechanisms of follicle-stimulating hormone (FSH) and Sphingosine-1-phosphate (S1P) on vitrified and post-transplantation ovaries. Ovaries from 21-day-old mice were cryopreservation by vitrification with 0.3 IU/mL FSH, 2 µM S1P, and 0.3 IU/mL FSH + 2 µM S1P, respectively, for follicle counting and detection of apoptosis-related indicators. The results demonstrated that FSH and S1P co-intervention during the vitrification process could preserve the primordial follicle pool and inhibit follicular atresia by suppressing cell apoptosis. The thawed ovaries were transplanted under the renal capsule of 6-8 week-old ovariectomized mice and removed 24 h or 7 days after transplantation. The results indicated that FSH and S1P co-intervention can inhibit apoptosis and autophagy in ovaries at 24 h after transplantation, and promote follicle survival by up-regulating Cx37 and Cx43 expression, enhanced angiogenesis in transplanted ovaries by promoting VEGF expression, as well as increased the E2 levels to restore ovarian endocrine function at 7 days after transplantation. The hypoxia and ischemia cell model was established by CoCl2 treatment for hypoxia in human granulosa-like tumor cell line (KGN), as well as serum-free culture system was used for ischemia. The results confirmed that ischemia-hypoxia-induced apoptosis in ovarian granulosa cells was reduced by FSH and S1P co-intervention, and granulosa cell autophagy was inhibited by up-regulating the AKT/mTOR signaling pathway. In summary, co-administration of FSH and S1P can maintain ovarian survival during ovarian vitrification and increase follicle survival and angiogenesis after transplantation.© 2023. BioMed Central Ltd., part of Springer Nature.