Introduction: A great attention is today given to the use of immature oocytes as alternative source of oocytes for assisted reproductive technologies (ARTs) [1]. Germinal Vesicle (GV) stage oocytes may be an important reserve (ARTs) for in vitro maturation (IVM) and improving cryopreservation quality [2]. This study aims to determine the ultrastructure of fresh GVs, taking in consideration the morphology of well-preserved organelles as indicator of quality post-IVM and post-cryopreservation. Methods: Fresh GV-stage oocytes were recovered from consenting donor women undergoing ARTs due to tubal factors. Ovarian stimulation was achieved with a long gonadotropin releasing hormone agonist (GnRH-A) protocol in combination with rFSH (150 IU/day). Oocyte retrieval was performed 34–36 h after ovulation, induced by 10000 IU of hCG. Oocytes were fixed and processed for standard preparative for transmission electron microscopy (TEM) observations [3,4]. Results: Light microscopy showed that all fresh GVs were of good quality. Ultrastructural data by TEM showed a roundish nucleus, numerous mitochondria often located in perinuclear position and sometimes associated to small vesicles (mitochondria-vesicle complexes), abundant vacuoles close to the nucleus, cortical granules dispersed throughout the whole ooplasm an intact and continuous zona pellucida and a physiological distribution of small or long microvilli, even if sometime uncontinous. Conclusion: These data on fresh human immature oocytes can be useful to evaluate the quality of hormonal stimulation and IVM protocols as well as ultrastructural damages after cryopreservation.
Ultrastructural analysis of human germinal vesicle-stage oocytes during application of assisted reproductive technologies / Antonouli, S.; Belli, M.; Palmerini, M. G.; Bianchi, S.; Bernardi, S.; Cecconi, S.; Donfrancesco, O.; Familiari, G.; Nottola, S. A.; Macchiarelli, G.. - (2018), pp. 4-4. (Intervento presentato al convegno XXVI International Symposium of Morphological Sciences tenutosi a Prague, Czech Republic).
Ultrastructural analysis of human germinal vesicle-stage oocytes during application of assisted reproductive technologies
O. Donfrancesco;G. Familiari;S. A. Nottola;
2018
Abstract
Introduction: A great attention is today given to the use of immature oocytes as alternative source of oocytes for assisted reproductive technologies (ARTs) [1]. Germinal Vesicle (GV) stage oocytes may be an important reserve (ARTs) for in vitro maturation (IVM) and improving cryopreservation quality [2]. This study aims to determine the ultrastructure of fresh GVs, taking in consideration the morphology of well-preserved organelles as indicator of quality post-IVM and post-cryopreservation. Methods: Fresh GV-stage oocytes were recovered from consenting donor women undergoing ARTs due to tubal factors. Ovarian stimulation was achieved with a long gonadotropin releasing hormone agonist (GnRH-A) protocol in combination with rFSH (150 IU/day). Oocyte retrieval was performed 34–36 h after ovulation, induced by 10000 IU of hCG. Oocytes were fixed and processed for standard preparative for transmission electron microscopy (TEM) observations [3,4]. Results: Light microscopy showed that all fresh GVs were of good quality. Ultrastructural data by TEM showed a roundish nucleus, numerous mitochondria often located in perinuclear position and sometimes associated to small vesicles (mitochondria-vesicle complexes), abundant vacuoles close to the nucleus, cortical granules dispersed throughout the whole ooplasm an intact and continuous zona pellucida and a physiological distribution of small or long microvilli, even if sometime uncontinous. Conclusion: These data on fresh human immature oocytes can be useful to evaluate the quality of hormonal stimulation and IVM protocols as well as ultrastructural damages after cryopreservation.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
