Ovary and ovulation: Follicular development in cryopreserved marmoset ovarian tissue after transplantation

Abstract

Pieces of marmoset ovary were frozen by slow cooling in 1.5 M dimethylsulphoxide. The follicles in fresh and frozen tissue were counted and examined for morphological appearance in stained serial sections. The proportion of normal follicles was similar in fresh tissue and frozen tissue examined immediately after thawing. Follicles at all stages of folliculogenesis up to the small antral stage survived freezing and thawing. Fresh and frozen tissue was transplanted underneath the kidney capsules of ovariectomized immunodeficient mice. The establishment of grafts was similar, and oestrogenic activity (cornification of the vaginal epithelium) was observed in the recipients 20 and 16 days after transplantation of fresh and frozen grafts respectively. The total number of follicles and the proportion of normal follicles were similar in fresh and frozen grafts. Grafts of frozen tissue recovered between 7 and 15 days after transfer contained follicles up to the small antral stage of development. Grafts recovered between 21 and 32 days contained follicles at all stages of folliculogenesis, including large antral follicles (1-2 mm diameter). Our results suggest that freezing and thawing do not substantially damage marmoset ovarian tissue, and the cryopreserved tissue retains its ability to support the development of large antral follicles.

Cryopreservation of caprine ovarian tissue using glycerol and ethylene glycol

“…The follicular degeneration (nuclear pyknosis and oocyte shrinkage) detected in the present study were also described by other authors after cryopreservation of human [14] and non-human primate ovarian tissue [11]. Additionally, the degeneration in the granulosa cell layer (detachment and disorganization) observed in our study has also been detected after vitrification of murine [13] and slow freezing of bovine [15] ovarian tissue.…”

“…The vials were plunged directly into liquid nitrogen (−196 °C) and stored for up to 5 days before thawing. The freezing curve used in this study was chosen because most procedures currently used to cryopreserve oocytes [25], [26], [27], [28], [29] and [30] and ovarian tissue [10], [11], [12], [14], [15], [18], [24], [31], [32] and [33] stipulate a cooling rate of 0.3-0.5 °C/min from the seeding temperature (usually −5 to −9 °C) to a lower temperature, usually between −30 and −40 °C.…”

“…Although there has been some success in ovarian tissue cryopreservation, several studies [11], [12], [13], [14] and [15] indicated that follicular damage occurs after freezing and thawing. Such damage can occur during any of step of the cryopreservation process (exposure of the tissue to the cryoprotectant, freezing or thawing).…”

Human oocyte and ovarian tissue cryopreservation and its application

“…Histological analysis indicated that a proportion of follicles are damaged during the freezing and thawing process in mouse, marmoset, and human ovarian tissue [90,98,99]. Nuclear and DNA damage in human and ovine follicles after freezing and thawing has been reported using immunohistochemistry, fluorescent in situ hybridization, and terminal deoxynucleotidyl transferase end labeling [100][101][102].…”

Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol

“…animals, including sheep [1], goats [2,3], primates [4], rodents [5], and humans [6]. Satisfactory results have been reported, especially when a 1.5 M concentration was used in slow-freezing protocols.…”