P-23: The Effect of LIF on In Vitro Growth and Apoptosis Incidence in Vitrified Human Ovarian Tissue after 14 days culture
The high effectiveness of vitrification, as a cryopreservation protocol, for human oocytes and embryos is shown, whereas data on human ovarian tissue are limited. This study aimed to assessment of follicular growth, ultrastructure, and apoptosis incidence in human ovarian tissue following vitrification/warming and after culture in the presence of LIF.
Materials and methods
Ovarian cortex biopsies from normal pregnant women fragmented and then divided to 2 main groups: vitrified and non-vitrified. Ovarian fragments in each main group cultured in 3 groups: in presence GDF-9B or LIF or absence any factor. Then the morphology, ultrastructure and incidence of apoptosis using biochemical analyses and analysis of apoptosis related genes expression in ovarian tissue fragments in all 6 experimental groups were evaluated before and after 2 weeks culture.
Morphology and ultrastructure of vitrified human ovarian tissue were better preserved in vitrified group in presence of LIF compared to GDF-treated and non-treated groups. 54 Int J Fertil Steril, Vol 9, Suppl 1, Summer 2015 Abstracts of The 16th Royan International Congress on Reproductive Biomedicine Apoptosis biochemical evaluations in all non-vitrified and vitrified treated groups were same. Apoptotic genes expression in both non-vitrified and vitrified groups at the end of culture period showed significant differences compared to non-treated groups. Morphological studies of ovarian tissue in all groups showed better conservation of ovarian follicles in presence of GDF- 9B and LIF treated groups (P<0.05). But there were no significant differences between non-vitrified and vitrified ovarian tissue in all treated groups. The levels of hormones at the end of culture period in treated groups were higher compared to non-treated groups. Apoptosis evaluation techniques showed that apoptosis incidence in treated groups were lower than non-treated cultured groups and non-cultured ovarian tissue (P<0.05).
This study showed that culture of vitrified human ovarian tissue has not increased the incidence of apoptosis and GDF-9B and LIF could improve survival and development of cultured vitrified ovarian fragments and reduce incidence of apoptosis in human ovarian tissue.