Current Issue

Volume 13, Number 4, Jan-Mar 2020 Pages: 330-338

Effects of Alginate Concentration and Ovarian Cells on In Vitro Development of Mouse Preantral Follicles: A Factorial Study


Parisa Jamalzaei, Ph.D, 1, Mojtaba Rezazadeh Valojerdi, Ph.D, 1, 2, *, Leila Montazeri, Ph.D, 3, Hossein Baharvand, Ph.D, 4, 5, *,
Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Department of Cell Engineering, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
Department of Developmental Biology, University of Science and Culture, Tehran, Iran
*Corresponding Addresses: P.O.Box: 14115-111 Department of Anatomy Faculty of Medical Sciences Tarbiat Modares University 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: 16635-148 Department of Stem Cells and Developmental Biology Cell Science Research Center Royan Institute for Stem Cell Biology and Technology ACECR Tehran Iran P.O.Box: 13145-871 Department of Developmental Biology University of Science and Culture Tehran Iran Emails:mr_valojerdi@modares.ac.ir,Baharvand@Royaninstitute.orgRoyan Institute

Abstract

Background

In the present study, the effects of alginate (ALG) concentration and ovarian cells (OCs) on the devel- opment and function of follicles were simultaneously evaluated.

Materials and Methods

In the first step of this experimental study, preantral follicles were isolated from the ovaries of 2-week-old mice, encapsulated in the absence or presence of OCs in 0.5, 0.75 and 1% ALG hydrogels, and cultured for 14 days. The morphology, diameter, survival and antrum formation rates of the follicles and the maturation of the oocytes were evaluated during culture. In the second step, preantral follicles were cultured in the best chosen ALG concentration, in both the absence and presence of OCs. Following these steps, the amount of DNA fragmentation, the expression levels of connexin 37 and connexin 43 proteins, the secretion levels of estradiol, progesterone and andros- tenedione by the follicles and the quality of mature (MII) oocytes were assessed.

Results

Our data revealed that in the absence of OCs, follicles of 0.5% group showed a higher survival rate than the 0.75 and 1% groups (71.87 vs. 52.52 and 40%, respectively, P<0.05). Nonetheless, the antrum formation rate of the 1% group was higher and its oocyte degeneration rate was lower than that in the other groups. Furthermore, it was observed that co-culture of follicles with OCs relatively increased the follicle diameter, survival, antrum formation, and germinal vesicle (GV) to GV break down (GVBD)/MII transition rates. At last, the comparison of 0.5%-OCs and 0.5%+OCs groups indicated that the co-culture condition resulted in more progesterone production (1.8 ± 0.2 vs. 3.2 ± 0.4 ng/ml, respectively, P<0.05) and also decreased oocytes’ cortical granule abnormalities (100 vs. 40% for 0.5%- OCs and 0.5%+OCs groups, respectively).

Conclusion

The present study revealed that 0.5% ALG hydrogel is relatively suitable for preantral follicle culture, and in the presence of OCs, it mimics the natural ovarian condition better than the higher concentrations of ALG hydrogel.