Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials Unit, Université Catholique de Louvain, Brussels, Belgium.
J Biomed Mater Res B Appl Biomater. 2022 May;110(5):1012-1022. doi: 10.1002/jbm.b.34974. Epub 2021 Nov 26.
To successfully assemble a bio-engineered ovary, we need to create a three-dimensional matrix able to accommodate isolated follicles and cells. The goal of this study was to develop an extracellular matrix hydrogel (oECM) derived from decellularized bovine ovaries able to support, in combination with alginate, human ovarian follicle survival and growth in vitro. Two different hydrogels (oECM1, oECM2) were produced and compared in terms of decellularization efficiency (dsDNA), ECM preservation (collagen and glycosaminoglycan levels), ultrastructure, rigidity, and cytotoxicity. oECM2 showed significantly less dsDNA, greater retention of glycosaminoglycans and better rigidity than oECM1. Isolated human ovarian follicles were then encapsulated in four selected hydrogel combinations: (1) 100% oECM2, (2) 90% oECM2 + 10% alginate, (3) 75% oECM2 + 25% alginate, and (4) 100% alginate. After 1 week of in vitro culture, follicle recovery rate, viability, and growth were analyzed. On day 7 of in vitro culture, follicle recovery rates were 0%, 23%, 65%, 82% in groups 1-4, respectively, rising proportionally with increased alginate content. However, there was no difference in follicle viability or growth between groups 2 and 3 and controls (group 4). In conclusion, since pure alginate cannot be used to graft preantral follicles due to its poor revascularization and degradation after grafting, oECM2 hydrogel combined with alginate may provide a new and promising alternative to graft isolated human follicles in a bio-engineered ovary.
为了成功组装生物工程卵巢，我们需要创建一个能够容纳分离的卵泡和细胞的三维基质。本研究的目的是开发一种源自去细胞牛卵巢的细胞外基质水凝胶（oECM），该水凝胶能够与藻酸盐结合，支持人卵巢卵泡在体外的存活和生长。我们制备了两种不同的水凝胶（oECM1、oECM2），并从去细胞效率（dsDNA）、细胞外基质保留（胶原蛋白和糖胺聚糖水平）、超微结构、刚性和细胞毒性方面对其进行了比较。oECM2 的 dsDNA 显著减少，糖胺聚糖保留更好，刚性更好。然后将分离的人卵巢卵泡包封在四种选定的水凝胶组合中：（1）100% oECM2；（2）90% oECM2+10%藻酸盐；（3）75% oECM2+25%藻酸盐；（4）100%藻酸盐。体外培养 1 周后，分析卵泡回收率、活力和生长情况。体外培养第 7 天，各组卵泡回收率分别为 0%、23%、65%、82%，随着藻酸盐含量的增加而呈比例上升。然而，在第 2 组和第 3 组与对照组（第 4 组）之间，卵泡活力或生长没有差异。总之，由于纯藻酸盐在移植后血管生成和降解不良，不能用于移植窦前卵泡，因此 oECM2 水凝胶与藻酸盐结合可能为移植分离的人卵泡提供一种新的、有前途的生物工程卵巢替代方法。
