In Vitro Development of Mouse Preantral Follicle with Using Amniotic Membrane Extract-Loaded Hydrogels

Abstract

An artificial ovary based on the alginate (ALG) hydrogel has been widely implemented to preserve prepubertal femalefertility. However, this platform is not fully capable of successful an ovary microenvironment simulation for follicledevelopment, holding great potential for its improvement. Therefore, this experimental study aimed to evaluate the effectof an amniotic membrane extract (AME) -loaded hydrogel on the mouse preantral follicles in vitro development.In order to have better follicle development, first, the impact of different concentrations of follicle-stimulating hormone(FSH) was evaluated on the mouse preantral follicles encapsulated in ALG. Later, the appropriate dose was adjustedfor the follicles encapsulated in the ALG-AME hydrogel. Results demonstrated that 100 mIU/ml FSH showeda significant follicle survival rate compared with 10 mIU/ml FSH (P=0.005). According to MTT assay finding, therate of weight loss, and rheology evaluations, ALG containing 1 mg/ml AME was identified as an optimal sampleof follicle culture instead of other AME concentrations. Follicle diameter significantly increased in the ALG-AME 1hydrogel compared with the ALG control group without AME (P=0.027). The storage modulus of ALG-AME 1 was773 Pa and retained the follicle morphology for 13 days. No statistically substantial difference was seen in survival,antrum cavity formation, and competent oocyte in terms of the normal chromosomal arrangement and meiotic spindlerate in comparison with the control group. It can be concluded that ALG-AME 1 could not significantly impact themouse preantral follicle.

Keywords

• alginate
• amniotic membrane extract
• preantral follicle
• hydrogel
• artificial ovary