438298Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
48499Department of Developmental Biology, University of Science and Culture, Tehran, Iran.
J Biomater Appl. 2022 Sep;37(3):563-574. doi: 10.1177/08853282221094193. Epub 2022 Apr 22.
Three-dimensional cultures of follicles on ECM-based scaffolds can be an approach for women who become infertile after cancer treatments. Human amniotic membrane (HAM) is extensively employed in tissue engineering because of its unique properties. We cultured mouse pre-antral follicles in a hydrogel derived from decellularized amniotic membrane (DAM) combined with alginate (ALG) to improve ovarian follicle culture. HAM was decellularized. Quantitative (nuclear contents, collagen, glycosaminoglycan [GAG]) and qualitative (DAPI, H&E, Masson's trichrome, Alcian blue, scanning electron microscopy assessments were performed. Then, we created an amniotic membrane-based hydrogel (AMBH) and conducted AMBH characterization assays (rheology, MTS, degradation rate). Isolated mouse pre-antral follicles were cultured in 15 mg/mL AMBH (AMBH15), 30 mg/mL AMBH (AMBH30), or 45 mg/mL AMBH (AMBH45). ALG hydrogel was the control group. Follicular diameters, estradiol hormone secretion rate, follicular morphology, and the follicle antral and degeneration rate were examined. Quantitative and qualitative assays indicated successful decellularization. AMBH characterization assays showed that the ALG hydrogel had more appropriate gelation and slower degradation than AMBH. There was a statistically higher antral follicle formation rate in the AMBH45 group ( < .05) compared to the AMBH30 and AMBH15 groups and less ( < .05) degenerated follicles. There was no significant difference with the ALG group. Diameter and estradiol hormone secretion in the AMBH45 group were not significantly higher than the ALG group. Although decellularization was confirmed and the viscoelastic parameters of AMBH support follicle culture, there was no significant effect on ovarian follicle maturation compared to the ALG control group.
基于细胞外基质支架的卵泡三维培养可能是癌症治疗后不孕女性的一种方法。人羊膜在组织工程中得到广泛应用，因为它具有独特的性质。我们将小鼠原始卵泡培养在脱细胞羊膜（DAM）与藻酸盐（ALG）结合衍生的水凝胶中，以改善卵巢卵泡培养。对羊膜进行脱细胞处理。进行了定量（核含量、胶原蛋白、糖胺聚糖[GAG]）和定性（DAPI、H&E、Masson 三色、阿利新蓝、扫描电子显微镜评估）检测。然后，我们创建了一种基于羊膜的水凝胶（AMBH）并进行了 AMBH 特性分析（流变学、MTS、降解率）。分离的小鼠原始卵泡在 15 mg/mL AMBH（AMBH15）、30 mg/mL AMBH（AMBH30）或 45 mg/mL AMBH（AMBH45）中培养。ALG 水凝胶为对照组。检查了卵泡直径、雌二醇激素分泌率、卵泡形态以及卵泡腔和退化率。定量和定性检测表明成功脱细胞。AMBH 特性分析表明，ALG 水凝胶的胶凝速度更快，降解速度更慢。与 AMBH30 和 AMBH15 组相比，AMBH45 组的腔前卵泡形成率更高（<0.05），退化卵泡更少（<0.05）。与 ALG 组无显著差异。AMBH45 组的直径和雌二醇激素分泌量与 ALG 组无显著差异。尽管确认了脱细胞处理，并且 AMBH 的黏弹性参数支持卵泡培养，但与 ALG 对照组相比，对卵巢卵泡成熟没有显著影响。
