Chitosan Hydrogel Supports Integrity Of Ovarian Follicles During In Vitro Culture: A Preliminary Of A Novel Biomaterial For Three Dimensional Culture Of Ovarian Follicles
Authors: Fatemeh Hassani, Bita Ebrahimi, Ashraf Moini, Ali Ghiaseddin, Mahshid Bazrafkan, Gholamreza Hassanzadeh, Mojtaba Rezazadeh Valojerdi
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Objective: Testing novel biomaterials for the three dimensional (3D) culture of ovarian follicles may ultimately lead to a culture
model which can support the integrity of follicles during in vitro culture (IVC). The present study reports the first application of
a chitosan (CS) hydrogel in culturing mouse preantral follicles.
Materials and Methods: In this interventional experiment study, CS hydrogels with the concentrations of 0.5, 1, and
1.5% were first tested for fourier transform infrared spectroscopy (FT-IR), Compressive Strength, viscosity, degradation,
swelling ratio, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity and live/dead assay.
Thereafter, mouse ovarian follicles were encapsulated in optimum concentration of CS (1%) and compared with those
in alginate hydrogel. The follicular morphology, quality of matured oocyte and steroid secretion in both CS and alginate
were assessed by enzyme-linked immunosorbent assay (ELISA). The expression of folliculogenesis, endocrine,
and apoptotic related genes was also evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) and
compared with day that in 0.
Results: The rates of survival, and diameter of the follicles, secretion of estradiol, normal appearance of meiotic
spindle and chromosome alignment were all higher in CS group compared with those in alginate group (P≤0.05). The
expression of Cyp19a1 and Lhcgr in CS group was significantly higher than that of the alginate group (P≤0.05).
Conclusion: The results showed that CS is a permissive hydrogel and has a beneficial effect on encapsulation of
ovarian follicle and its further development during 3D culture.