Agar; A Suitable Scaffold for Culturing of Human Ovarian Tissue

Background: Nowadays ovarian tissue banks have been set up in many countries to improve chances of child bearing for cancer patients. As transplantation of cryopreserved ovary in-tensifies the possibility of malignant cells reintroduction, re-searchers are focusing more on ovarian tissue in-vitro culture methods. By tissue engineering and biomaterial improvement, embryologists attempt to use various scaffolds to mimic the ovarian extracellular matrix and preserve the tissue structure in the culture period. Our aim was to achieve an optimum culture system for human ovarian tissue to activate higher numbers of primordial follicles.Materials and Methods: Donated ovarian tissues from 6 trans-sexual women were dissected, cryopreserved by slow-freezing method and cultured on either matrigel coated insert or agar-soaked scaffold for 7 days. The numbers of morphologically healthy and degenerated follicles, their developmental stages, the hormonal levels of the spent medium and also the relative expression of the apoptotic pathway and proliferation genes were evaluated and compared.Results: Significant decrease in the number of primordial fol-licles was observed in both cultured groups in comparison to non-cultured control group. Besides, the number of degener-ated follicles was higher in both cultured groups than the nonculture control group, but it was only significant for primary follicles in insert cultured group. The number of primary and secondary healthy follicles was significantly higher in the agar cultured ovarian strips compared to the non-cultured control group. A significant increase was seen in estrogen level in the agar cultured group at day 7 when compared to the second day of culture. The Q-PCR analysis showed a down regulation of apoptotic genes and an increase in Ki67 gene during culture in both groups compared to the non-cultured control group.Conclusion: Since there was no significant difference between these two scaffolds in evaluated parameters and on the other hand there was a significant follicular growth in agar cultured group in comparison to non-cultured control; agar scaffold can be recommended as a suitable scaffold for human ovarian tissue culture. Meanwhile it must be mentioned that agar is less ex-pensive and more accessible compared to other scaffolds used in this field.

Keywords:

In Vitro Culture , Cryopreservation , Human Ovarian Tissue , Agar , Scaffold