Oxygen consumption rate of early pre-antral follicles from vitrified human ovarian cortical tissue.

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.

Evaluation of oxygen consumption in human vitrified and warmed pre-antral follicles after prolonged low temperatures.

PURPOSE: The purpose of this study was to evaluate the effect of transportation at prolonged low temperatures on the survival of pre-antral follicles. METHODS: Ovarian tissue was removed from six women with gender identity disorder. Tissues were stored in an icebox at 4 °C for 6 or 18 h prior to vitrification. After warming, ovarian tissues were cultured for 24 h and follicle survival was assessed via a viability/cytotoxicity kit. Morphological features and oxygen consumption rate (OCR) were evaluated by scanning electrochemical microscopy (SECM). RESULTS: Survival rate of isolated primordial follicles was 95.7 and 100 %, and that of primary follicles was 91.7 and 81.8 % in the 6- and 18-h groups respectively. There was no difference in morphology between the 6- and 18-h storage groups. In comparison with OCR of vitrified-warmed follicles and OCR of 24-h culture after vitrified-warmed follicles, OCR of 24-h culture after vitrified-warmed primordial follicles was significantly higher in both 6-hour (0.02 ± 0.02 vs 0.07 ± 0.04, P < 0.05) and 18-h groups (0.02 ± 0.02 vs 0.11 ± 0.10, P < 0.05). CONCLUSIONS: This strongly suggests that prolonged transportation of ovarian tissue at low temperatures is useful when there are no available local systems for fertility preservation.

A birth from the transfer of a single vitrified-warmed blastocyst using intracytoplasmic sperm injection with calcium ionophore oocyte activation in a globozoospermic patient.

OBJECTIVE: To present the effectiveness of diagnostic heterologous intracytoplasmic sperm injection (ICSI), mouse oocyte activation test (MOAT), and ICSI combined with assisted oocyte activation (AOA) in a globozoospermic patient. DESIGN: A case report. SETTING: A private IVF center, Japan. PATIENT(S): A patient with globozoospermia. INTERVENTION(S): MOAT in a mouse and ICSI combined with AOA in a human. MAIN OUTCOME MEASURE(S): Ultrastructure, MOAT, fertilization, and pregnancy. RESULT(S): The transmission electron micrographs showed 100% round-headed spermatozoa lacking an acrosome. MOAT showed that the fertilization rate was 68.4% (13/19) when AOA was used but 0% (0/19) when AOA was not used. After the diagnosis of globozoospermia and sperm-related activation deficiency, 17 human mature oocytes were activated with calcium ionophore after ICSI was performed. The fertilization rate was 88.2% (15/17), and 11 blastocysts were cryopreserved using the vitrification method to prevent severe ovarian hyperstimulation syndrome. A single vitrified-warmed blastocyst was transferred. A gestational sac with fetal heart movements was recognized, and a healthy boy weighing 3180 g was born at 40 weeks of gestation by cesarean section without any congenital abnormality. CONCLUSION(S): MOAT allows discrimination between sperm- and oocyte-related fertilization failures and shows the effectiveness of AOA.