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Oxygen consumption rate of early pre-antral follicles from vitrified human ovarian cortical tissue
Authors:Takayuki ISHIKAWA  Toshihiko KYOYA  Yusuke NAKAMURA  Eimei SATO  Tatsuhiro TOMIYAMA  Koichi KYONO
Institution:1)Reproduction Research Center, Kyono ART Clinic, Sendai 980-0014, Japan;2)Graduate School of Agricultural Science, Tohoku University, Sendai 981-8555, Japan;3)New ART Research Center, Osaka New ART Clinic, Osaka 530-0001, Japan
Abstract:The study of human ovarian tissue transplantation and cryopreservation has advanced significantly. Autotransplantation of human pre-antral follicles isolated from cryopreserved cortical tissue is a promising option for the preservation of fertility in young cancer patients. The purpose of the present study was to reveal the effect of vitrification after low-temperature transportation of human pre-antral follicles by using the oxygen consumption rate (OCR). Cortical tissues from 9 ovaries of female-to-male transsexuals were vitrified after transportation (6 or 18 h). The follicles were enzymatically isolated from nonvitrified tissue (group I, 18 h of transportation), vitrified-warmed tissue (group II, 6 and 18 h of transportation) and vitrified-warmed tissue that had been incubated for 24 h (group III, 6 and 18 h of transportation). OCR measurement and the LIVE/DEAD viability assay were performed. Despite the ischemic condition, the isolated pre-antral follicles in group I consumed oxygen, and the mean OCRs increased with developmental stage. Neither the transportation time nor patient age seemed to affect the OCR in this group. Meanwhile, the mean OCR was significantly lower (P < 0.05) in group II but was comparable to that of group I after 24 h of incubation. The integrity of vitrified-warmed primordial and primary follicles was clearly corroborated by the LIVE/DEAD viability assay. These results demonstrate that the OCR can be used to directly estimate the effect of vitrification on the viability of primordial and primary follicles and to select the viable primordial and primary follicles from vitrified-warmed follicles.
Keywords:Human ovarian cortical tissue  Isolation  Oxygen consumption rate  Pre-antral follicle  Vitrification
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