Histochemical and Immunohistochemical Investigation of the Effects of Cryoprotectants and Cryopreservation with PRP-Added Vitrification Solution on Rat Ovarian Tissue
Project no. 20518 · Status: Kapanmış · 2019
Summary
In situations that may cause infertility in women, such as radiotherapy and chemotherapy applications used in cancer treatments, some drugs used to suppress the immune system and surgical interventions used to treat gynecological disorders, there is a need for treatment options to preserve fertility. Oocyte, embryo or ovarian tissue can be cryopreserved and stored in order to maintain fertility. Although oocyte and embryo storage is routinely practiced in assisted reproductive centers, ovarian tissue cryopreservation is not a standard practice method accepted by everyone, thus it has not been used in routine clinical use yet and has remained at an experimental level. Experimental studies on this subject continue in many national and foreign research centers. Within this study, which is the subject of the research project, 5 study groups with 7 experimental animals in each group were formed. Group 1 (n = 7): Control group; Group 2 (n = 7): The group that was followed up directly after cryopreservation with vitrification solution containing PRP; Group 3 (n = 7): The group that is stored in liquid nitrogen at -196oC for 6 months after cryopreservation process with the PRP containing vitrification solution; Group 4 (n = 7): The group that was followed up directly after cryopreservation with a PRPfree vitrification solution; Group 5 (n = 7): The group that is stored in liquid nitrogen at -196 oC for 6 months after cryopreservation with a PRP-free vitrification solution. In the preparation of vitrification solutions, Ethylene Glycol (EG) and dimethylsulfoxide (DMSO) were used as cryoprotectants. PRP obtained after centrifugation of 4 ml of blood taken by cardiac puncture before oophorectomy from experimental animals under deep anesthesia in Group 2 and Group 3 was activated with 10% CaCl2 and used in the preparation of vitrification solution applied in Group 2 and Group 3. In this study, histochemical and immunohistochemical analysis of the acute and long-term effects of cryoprotectants and vitrification solutions containing PRP used in cryopreservation on ovarian tissue cryopreservation was carried out to contribute to the standardization of the cryopreservation method to be applied and the solutions to be used. Keywords: Cryoprotectant, EG, DMSO, PRP, vitrification, ovarian tissue cryopreservation