Vatanparast Mahboubeh, Maleki Behnam, Khalili Mohammad Ali
Molecular Medicine Research Center, Research Institute of Basic Medical Sciences, Rafsanjan University of Medical Sciences, Rafsanjan, Iran.
Department of Reproductive Biology, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran; Infertility Center, Department of Obstetrics and Gynecology, Mazandaran University of Medical Sciences, Sari, Iran.
Eur J Obstet Gynecol Reprod Biol. 2023 May;284:94-99. doi: 10.1016/j.ejogrb.2023.03.013. Epub 2023 Mar 15.
The first days of post-ovarian transplantation are critical periods, as the ischemic injury can diminish the success rate. In this study, the first day's events of ovarian transplantation in two dimensions of structure and ultrastructure following slow freezing and vitrification were assessed.
Ovarian tissues (OTs) from 10 cancerous patients were frozen in two methods of slow freezing and vitrification. Tissues were transplanted onto the CAM and then retrieved at 5 and 10 days of culture. Nine groups were assigned as follows; I-III; fresh, 5 and 10 days culture, IV-VI; vitrification, 5 and 10 days culture, and VII-IX; slow freezing, 5 and 10 days culture. Structural and ultra-structural studies were done to assess the tissue viability and integrity following CAM transplantation. Image J software was used to measure the amounts of fibrosis and necrosis.
The first sign of successful transplantation was found on day 3 post-transplantation. Vitrified tissues showed higher viability and transplantation rate compared to the slow frozen group (65% vs 57.5%) (p = 0.7). Tissue fibrosis and areas didn't increase significantly after cryopreservation using two methods (p > 0.05). The areas of fibrosis and necrosis and avian vessels increased significantly after 5 and 10 days of culture (p < 0.05). Large ultra-structural follicular deformities were noticed after 10 days of CAM transplantation. Better stromal ultrastructure features can be found after vitrified tissue culture. Also, the CAM transplantation technique had negative effects on the integrity of follicles, independent of the freezing procedure.
Evaluation of early events of the ovarian post-transplantation is of amount importance, since the hypoxia during this period may accelerate follicular pool depletion, before the tissue stability. Vitrification can be considered a reliable alternative for slow freezing. CAM transplantation is a good technique for confirmation of tissue viability after warming but damaged the follicle ultrastructure in a short period.
卵巢移植后的最初几天是关键时期，因为缺血性损伤会降低成功率。在本研究中，评估了慢速冷冻和玻璃化冷冻后卵巢移植第一天在结构和超微结构两个维度上的情况。
将10例癌症患者的卵巢组织（OTs）分别采用慢速冷冻和玻璃化冷冻两种方法进行冷冻。将组织移植到鸡胚绒毛尿囊膜（CAM）上，然后在培养5天和10天时取出。分为九组如下：I - III组，新鲜组织、培养5天和10天的组织；IV - VI组，玻璃化冷冻组织、培养5天和10天的组织；VII - IX组，慢速冷冻组织、培养5天和10天的组织。进行结构和超微结构研究以评估CAM移植后组织的活力和完整性。使用Image J软件测量纤维化和坏死的程度。
移植后第3天发现成功移植的首个迹象。与慢速冷冻组相比，玻璃化冷冻组织显示出更高的活力和移植率（65%对57.5%）（p = 0.7）。使用两种方法冷冻保存后，组织纤维化和面积没有显著增加（p > 0.05）。培养5天和10天后，纤维化、坏死区域及鸡胚血管面积显著增加（p < 0.05）。CAM移植10天后观察到大量超微结构卵泡畸形。玻璃化冷冻组织培养后可发现更好的基质超微结构特征。此外，CAM移植技术对卵泡完整性有负面影响，与冷冻程序无关。
评估卵巢移植后的早期情况非常重要，因为在此期间的缺氧可能在组织稳定之前加速卵泡池的消耗。玻璃化冷冻可被视为慢速冷冻的可靠替代方法。CAM移植是一种确认复温后组织活力的良好技术，但在短时间内会损害卵泡超微结构。
