Mouse ovarian follicle growth in an amniotic membrane-based hydrogel

Abstract

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 ( p < .05) compared to the AMBH30 and AMBH15 groups and less ( p < .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.

Extracellular matrix-derived scaffolds in constructing artificial ovaries for ovarian failure: a systematic methodological review

“…Two studies utilized tissues from humans and other species simultaneously ( Table 1 ). The most common source of dECM scaffolds was ovarian tissues (n = 8), followed by amniotic membranes (n = 2), greater omentum (n = 1), and peritoneal membranes (n = 1) ( Motamed et al , 2017 ; Sarabadani et al , 2021 ; Haghshenas et al , 2022 ). In seven studies, different regions of tissues (six ovaries and one amniotic membrane) were dissected and subsequently minced ( Supplementary Table S4 ) ( Laronda et al , 2015 ; Motamed et al , 2017 ; Hassanpour et al , 2018 ; Nikniaz et al , 2021 ; Chiti et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ).…”

“…In seven studies, different regions of tissues (six ovaries and one amniotic membrane) were dissected and subsequently minced ( Supplementary Table S4 ) ( Laronda et al , 2015 ; Motamed et al , 2017 ; Hassanpour et al , 2018 ; Nikniaz et al , 2021 ; Chiti et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ). The greater omentum and amniotic membrane were directly cut into small fragments ( Fazelian-Dehkordi et al , 2022 ; Haghshenas et al , 2022 ). Four studies conducted freeze–thaw cycles.…”

“…Seven studies used detergent combinations ( Nikniaz et al , 2021 ; Pennarossa et al , 2021 ; Sarabadani et al , 2021 ; Chiti et al , 2022 ; Fazelian-Dehkordi et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ). From the perspective of chemical types, ionic detergents (SDS, sodium deoxycholate, SLES) were the most commonly used reagents (n = 10) ( Laronda et al , 2015 ; Hassanpour et al , 2018 ; Alaee et al , 2021 ; Nikniaz et al , 2021 ; Pennarossa et al , 2021 ; Chiti et al , 2022 ; Haghshenas et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ), and five studies used non-ionic detergents, mainly Triton X-100 ( Nikniaz et al , 2021 ; Pennarossa et al , 2021 ; Chiti et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ). Less frequently used detergents included ammonium hydroxide and 2-propanol.…”

“…Trypsin/EDTA and DNase/RNase solution were equally frequently used enzymes following detergents (n = 6) ( Motamed et al , 2017 ; Hassanpour et al , 2018 ; Sarabadani et al , 2021 ; Chiti et al , 2022 ; Zheng et al , 2022 ; Wu et al , 2022b ). In three studies, the dECM scaffolds were further processed into hydrogels, which were easier to use and apply ( Supplementary Table S4 ) ( Chiti et al , 2022 ; Haghshenas et al , 2022 ; Zheng et al , 2022 ). Regarding the operation duration, most studies completed decellularization within several days, except for those that used intact ovaries, where the operation spanned >3 weeks ( Laronda et al , 2015 ; Hassanpour et al , 2018 ).…”

Construction of Artificial Ovaries with Decellularized Porcine Scaffold and Its Elicited Immune Response after Xenotransplantation in Mice

“…Table S1: Quantification comparisons between different ovarian decellularization protocols. References [ 45 , 46 , 47 ] are cited in the supplementary materials.…”