Chitosan Hydrogel Supports Integrity of Ovarian Follicles during In Vitro Culture: A Preliminary of A Novel Biomaterial for Three Dimensional Culture of Ovarian Follicles


Fatemeh Hassani, M.Sc, 1Bita Ebrahimi, Ph.D, 2Ashraf Moini, Ph.D, 3,4Ali Ghiaseddin, Ph.D, 5Mahshid Bazrafkan, Ph.D, 1Gholamreza Hassanzadeh, Ph.D., 1,*Mojtaba Rezazadeh Valojerdi, Ph.D., 2,6,*
Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Department of Gynecology and Obstetrics, Roointan-Arash Maternity Hospital, Tehran University of Medical Sciences, Tehran, Iran
Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Biomedical Engineering Group, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, Iran
Department of Anatomy, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran
Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Department of Gynecology and Obstetrics, Roointan-Arash Maternity Hospital, Tehran University of Medical Sciences, Tehran, Iran
Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Biomedical Engineering Group, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, Iran
Department of Anatomy, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran
*Corresponding Addresses: P.O.Box: 14115-111 Department of Anatomy School of Medicine Tehran University of Medical Sciences Tehran Iran P.O.Box: 16635-148 Department of Embryology Reproductive Biomedicine Research Center Royan Institute for Reproductive Biomedicine ACECR Tehran Iran P.O. Box: 14115-111 Department of Anatomy Faculty of Medical Science Tarbiat Modares University Tehran Iran Emails:hassanzadeh@tums.ac.ir,mr_valojerdi@royaninstitute.org,mr_valojerdi@modares.ac.ir
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Hassani Fatemeh, Ebrahimi Bita, Moini Ashraf, Ghiaseddin Ali, Bazrafkan Mahshid, Hassanzadeh Gholamreza, Rezazadeh Valojerdi Mojtaba. Chitosan Hydrogel Supports Integrity of Ovarian Follicles during In Vitro Culture: A Preliminary of A Novel Biomaterial for Three Dimensional Culture of Ovarian Follicles . Cell J. 2020; 21(4): 479-493.

Abstract

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.