Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei, 430071, China.
J Assist Reprod Genet. 2020 Jul;37(7):1745-1753. doi: 10.1007/s10815-020-01797-4. Epub 2020 May 20.
This study was performed to investigate the effect of hypothermic machine perfusion (HMP) after cold storage (CS) on ovarian transplantation.
Rats aged 8-10 weeks were used as the donors and recipients for allotransplantation. Eighteen donor rats were divided into three groups: the fresh control (n = 6), cold storage (CS; n = 6), and hypothermic machine perfusion (HMP; n = 6) groups. The preservation solution contained Dulbecco's modified Eagle's medium/Ham's F-12 (1:1, v/v), 10% fetal bovine serum, 10 μg/ml insulin, 10 μg/ml transferrin, and 50 mIU/ml follicle-stimulating hormone (FSH). The donor ovaries in the CS and HMP groups were excised and then respectively subjected to 4 h of CS and 2 h of CS combined with 2 h of HMP at 4 °C, and then transplanted beneath the recipient's left renal capsule. At 7 days after transplantation, the ovaries were removed and blood samples were obtained for histological analysis, immunohistochemistry for CD31 and Ki67, and serum anti-Mullerian hormone (AMH) level estimation.
The HMP group showed significant increases in serum AMH and CD31-positive areas when compared to these values in the CS group (P < 0.05). However, no differences were noted in the total number of follicles or the Ki67-positive areas among the three groups.
Hypothermic machine perfusion after static cold storage is more effective than static CS alone for the short-term preservation of whole ovaries during transport. Whole ovary transplantation with vascular pedicle is our future research direction. Graphical Abstract The black rectangle in the figure shows the place where ligation and disconnection are required, the black dotted line shows the place where vascular forceps are used to clamp, and the black circle shows the place where the cannula is inserted This diagram was made for reviewers to understand more intuitively how my hypothermia mechanical perfusion model was built. Organs obtained in this way can be used for subsequent perfusion and whole ovarian transplantation.
本研究旨在探讨低温机器灌注（HMP）在冷保存（CS）后对卵巢移植的影响。
将 8-10 周龄大鼠作为同种异体移植的供体和受体。18 只供体大鼠分为三组：新鲜对照组（n = 6）、冷保存组（CS；n = 6）和低温机器灌注组（HMP；n = 6）。保存液含 Dulbecco's 改良 Eagle 培养基/Ham's F-12（1:1，v/v）、10%胎牛血清、10μg/ml 胰岛素、10μg/ml 转铁蛋白和 50mIU/ml 卵泡刺激素（FSH）。CS 组和 HMP 组的供体卵巢分别切除后，在 4°C 下分别进行 4 h CS 和 2 h CS 联合 2 h HMP，然后移植到受体左肾包膜下。移植后 7 天，取出卵巢并采集血液样本进行组织学分析、CD31 和 Ki67 的免疫组化分析以及血清抗苗勒管激素（AMH）水平测定。
与 CS 组相比，HMP 组血清 AMH 和 CD31 阳性面积显著增加（P < 0.05）。然而，三组之间总卵泡数或 Ki67 阳性面积无差异。
与单纯静态 CS 相比，低温机器灌注后静态 CS 更有利于运输过程中整个卵巢的短期保存。带血管蒂的全卵巢移植是我们未来的研究方向。
图中黑色矩形表示需要结扎和断开的部位，黑色虚线表示使用血管钳夹的部位，黑色圆圈表示插入套管的部位。此图是为了让审稿人更直观地了解我的低温机械灌注模型是如何构建的。通过这种方式获得的器官可用于后续灌注和全卵巢移植。
