Optimal vitrification protocol for mouse ovarian tissue cryopreservation: effect of cryoprotective agents and in vitro culture on vitrified-warmed ovarian tissue survival

Abstract

We compared eight vitrification protocols according to CPA composition and found the EDS protocol to be the optimal method among them. The data presented herein will help improve OT cryopreservation protocols for humans or other animals.

Effect of Exogenous Anti-Müllerian Hormone Treatment on Cryopreserved and Transplanted Mouse Ovaries

“…A follicle with over 30% apoptotic-positive cells, indicated by green fluorescence, was counted as an apoptotic follicle. 19 …”

“…15,16 For prevention of ischemic injury or cryodamage, many studies have been performed in different species. [17][18][19][20][21] Nevertheless, there have been no studies regarding prevention of massive follicular loss due to FSH overstimulation of primordial follicles.…”

Transplantation of mouse ovarian tissue: Comparison of the transplantation sites

“…Each follicle was classified into four types according to the categories of Lundy et al [23]; primordial (single layer of flattened pregranulosa cells), primary (single-layer granulosa cells [GC] including cuboidal forms), secondary (at least two layers of cuboidal GC), and antral (multiple layers of cuboidal GC with antrum). The integrity of each follicle was evaluated using the following criteria by Youm et al [24]; G1 (good: intact spherical follicle and oocyte), G2 (fair: GC pulled away from edge of follicles, but with intact oocyte), and G3 (poor: disruption and loss of granulosa and theca cells, pyknotic nuclei, and missing oocyte). To avoid miscounting, the follicles were analyzed in only one section per OT when they contained oocytes.…”

The beneficial effects of polyethylene glycol-superoxide dismutase on ovarian tissue culture and transplantation

“…Ovaries were collected from the ovarian bursa and transferred directly into vitrification media using the two-step method described by Youm et al [15]. First, the ovaries were equilibrated in 7.5 % ethylene glycol and 7.5 % dimethyl sulfoxide (DMSO) in 1× phosphate-buffered saline (PBS) supplemented with 20 % fetal bovine serum (FBS) for 10 min at room temperature (RT).…”