RESUMO
BACKGROUND: Intracellular lipids are sensitive to freezing. Lipidome modification is an important tool for studying the role of intracellular lipids in cryotolerance of mammalian oocytes and preimplantation embryos. OBJECTIVE: To study the effects of in vitro exposure of murine embryos to saturated stearic acid (SA) on the lipid content, embryo development and cryotolerance. MATERIALS AND METHODS: In vivo derived mouse embryos were cultured with 100 uM SA for 48 h up to the morula/blastocyst stage. Some of the SA-treated embryos were chosen for the evaluation of their development competence and the change in the lipidome, and other embryos were either slowly frozen or rapidly vitrified. RESULTS: Nile red staining combined with confocal laser scanning microscopy revealed a decrease in the total amount of lipids in the SA-treated embryos. Raman measurements showed that the lipid unsaturation was lower in embryos after in vitro SA culture. The addition of SA did not affect the embryo development before cryopreservation, but negatively affected the results of slow freezing cryopreservation and vitrification. CONCLUSION: In vitro SA exposure lowered the total amount of intracellular lipids and unsaturation in mouse embryos. The changes were accompanied with a significantly lower efficacy of embryo cryopreservation. https://doi.org/10.54680/fr24110110512.
Assuntos
Criopreservação , Ácidos Esteáricos , Vitrificação , Animais , Camundongos , Criopreservação/métodos , Embrião de Mamíferos , Blastocisto , Desenvolvimento Embrionário , Lipídeos , Técnicas de Cultura Embrionária , MamíferosRESUMO
Under physiological conditions, the membranes and lipid droplets of germ cells are in a conformationally disordered phase. Typically, during cooling, lipids undergo the transition to ordered phases and, upon heating, melt into a disordered phase. In this communication, we report the lipid phase transition in lipid droplets observed in porcine oocytes. Upon cooling, a sharp lipid phase transition from conformationally disordered to ordered state was detected within the temperature range between 20 and 15 °C. Subsequent heating to 45 °C does not return lipids to their original phase state. To the best of our knowledge, this is the first observation of an irreversible phase transition in lipid droplets of biological cells with native lipid composition.
Assuntos
Criopreservação , Oócitos , Animais , Suínos , Criopreservação/métodos , Transição de Fase , Congelamento , LipídeosRESUMO
Cryopreservation of preimplantation embryos is a widely used technique, but this procedure might impact the subsequent embryo development. The effect of slow freezing and vitrification on the lipid metabolism in preimplantation mammalian embryos is not well studied. In this work, we applied Raman spectroscopy of isotopically labeled molecules to address the effects of cryopreservation on fatty acid accumulation in mouse embryos. Embryos after slow freezing or vitrification were cultured for 20 h in a medium supplemented with bovine serum albumin saturated with deuterated stearic acid (dSA). After this period the concentration of dSA estimated from Raman spectra of frozen-thawed and vitrified-warmed embryos at the morula stage was almost twice higher compared to non-cryopreserved morulas. At the same time, frozen-thawed and vitrified-warmed 4-cell embryos did not demonstrate any difference in the level of stearic acid uptake from non-cryopreserved embryos of the same stage. After an additional 24 h culture, cryopreserved and non-cryopreserved embryos demonstrated similar dSA uptake.
Assuntos
Criopreservação , Transferência Embrionária , Camundongos , Animais , Criopreservação/métodos , Transferência Embrionária/métodos , Vitrificação , Blastocisto , MamíferosAssuntos
Ablação por Cateter/métodos , Histiocitoma Fibroso Benigno/cirurgia , Neoplasias/cirurgia , Baço/cirurgia , Esplenectomia/métodos , Neoplasias Esplênicas/cirurgia , Adulto , Ablação por Cateter/instrumentação , Hemostasia , Histiocitoma Fibroso Benigno/irrigação sanguínea , Histiocitoma Fibroso Benigno/patologia , Humanos , Masculino , Neoplasias/irrigação sanguínea , Neoplasias/patologia , Baço/irrigação sanguínea , Baço/patologia , Esplenectomia/instrumentação , Neoplasias Esplênicas/irrigação sanguínea , Neoplasias Esplênicas/patologia , Ultrassonografia/instrumentação , Ultrassonografia/métodosRESUMO
Biophysical peculiarities of action on tissues of a two-strem low-frequency ultrasound (TSLFU) technology, elaborated by "Arobella Medical LLC" (USA) firm, were studied. Capacity of ultrasound to separate a pathologically-changed and healthy tissues, to divide the structures in accordance to their bioacoustical parameters constitutes the technology peculiarities. The presence of such a biophysical effect permits to achieve high resectability (R0) in patients with oncological diseases. Antibacterial effect and stimulation of intraorgan microcirculation with ultrasound irradiation were noted. Biophysical peculiarities of TSLFU were successfully applied in surgical treatment of 48 patients, suffering inflammatory and oncological diseases of the abdominal cavity organs.