Division of Obstetrics and Gynecology, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Huddinge, 14186, Stockholm, Sweden.
Department of Gynecology and Reproductive Medicine, Karolinska University Hospital, 14186, Stockholm, Sweden.
J Assist Reprod Genet. 2024 Apr;41(4):1035-1055. doi: 10.1007/s10815-024-03038-4. Epub 2024 Feb 15.
Ovarian tissue cryopreservation is vital for fertility preservation, yet its effect on ovarian tissue follicle survival and transcriptomic signature requires further investigation. This study delves into the effects of vitrification on tissue morphology, function, and transcriptomic changes, helping to find possibilities for vitrification protocol improvements.
Ovarian cortex from 19 bovine animals were used to conduct pre- and post-vitrification culture followed by histological assessment, immunohistochemistry, and TUNEL assay. Follicles' functionality was assessed for viability and growth within the tissue and in isolated cultures. RNA-sequencing of ovarian tissue was used to explore the transcriptomic alterations caused by vitrification.
Follicle density, cell proliferation, and DNA damage in ovarian stroma were unaffected by vitrification. However, vitrified cultured tissue exhibited reduced follicle density of primordial/primary and antral follicles, while freshly cultured tissue manifested reduction of antral follicles. Increased stromal cell proliferation and DNA damage occurred in both groups post-culture. Isolated follicles from vitrified tissue exhibited similar viability to fresh follicles until day 4, after which the survival dropped. RNA-sequencing revealed minor effects of vitrification on transcriptomic signatures, while culture induced significant gene expression changes in both groups. The altered expression of WNT and hormonal regulation pathway genes post-vitrification suggests the molecular targets for vitrification protocol refinement.
Vitrification minimally affects tissue morphology, follicle density, and transcriptomic signature post-thawing. However, culture revealed notable changes in vitrified tissue samples, including reduced follicle density, decreased isolated follicle survival, and alteration in WNT signalling and ovarian hormonal regulation pathways, highlighted them as possible limitations of the current vitrification protocol.
卵巢组织冷冻保存对生育力保存至关重要，但它对卵巢组织卵泡存活和转录组特征的影响仍需进一步研究。本研究深入探讨了玻璃化对组织形态、功能和转录组变化的影响，有助于寻找改进玻璃化方案的可能性。
本研究使用 19 头奶牛的卵巢皮质进行玻璃化前后的培养，随后进行组织学评估、免疫组织化学和 TUNEL 检测。评估了卵泡在组织内和分离培养中的活力和生长情况，以评估其功能。使用卵巢组织的 RNA 测序来探索玻璃化引起的转录组改变。
玻璃化对卵巢基质中的卵泡密度、细胞增殖和 DNA 损伤没有影响。然而，玻璃化培养的组织表现出原始/初级和腔前卵泡的卵泡密度降低，而新鲜培养的组织表现出腔前卵泡减少。培养后两组基质细胞增殖和 DNA 损伤增加。来自玻璃化组织的分离卵泡在第 4 天之前与新鲜卵泡具有相似的活力，之后其存活率下降。RNA 测序显示玻璃化对转录组特征的影响较小，而培养在两组中均引起显著的基因表达变化。玻璃化后 WNT 和激素调节途径基因的表达改变表明了玻璃化方案改进的分子靶标。
玻璃化解冻后对组织形态、卵泡密度和转录组特征的影响最小。然而，培养揭示了玻璃化组织样本的显著变化，包括卵泡密度降低、分离卵泡存活率降低以及 WNT 信号和卵巢激素调节途径的改变，这些变化突出了当前玻璃化方案的潜在局限性。
