TY - JOUR

T1 - Alteration of the lipid phase transition during mouse embryos freezing after in vitro culture with linoleic acid

AU - Igonina, T. N.

AU - Okotrub, K. A.

AU - Brusentsev, E. Yu

AU - Chuyko, E. A.

AU - Ragaeva, D. S.

AU - Ranneva, S. V.

AU - Amstislavsky, S. Ya

N1 - Funding Information: This study was partly supported by the Russian Foundation for Basic Research (Project No. 19-016-00025 ) and by the budgeted project of the Institute of Cytology and Genetics (Novosibirsk, Russia) No. 0259-2021-0015 . Part of this work was supported by the budgeted project of the Institute of Automation and Electrometry (Novosibirsk, Russia) State Assignment No. AAAA-A17-117052410033-9 . The studies are implemented using the equipment of the Center for Genetic Resources of Laboratory Animals at ICG SB RAS, supported by the Ministry of Education and Science of Russia (Unique identifier of the project RFMEFI62119X0023 ). Part of the experiments was performed in the Multipleaccess center “High-resolution spectroscopy of gases and condensed matters” in IA&E SB RAS (Novosibirsk, Russia). Publisher Copyright: © 2021 Elsevier Inc. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/4

Y1 - 2021/4

N2 - Lipids significantly affect embryo cryopreservation in some mammalian species depending on the cell lipidome quantity and composition. One of the ways to study the relationship between lipid content and cryotolerance of cells is to study the effect of lipidome modification on laboratory mice. The objective of this research was to study how in vitro culture of mouse embryos with linoleic acid (LA) will affect lipid phase transition (LPT) during cooling and subsequent embryo development after cryopreservation. Embryos obtained in vivo at the 2-cell stage were cultured with 200 μM LA for 46 h up to the morula/blastocyst stage. Thereafter, one portion of embryos was slowly frozen to reveal the effect of LA on survival after cryopreservation, another portion was used to characterize the lipid composition and to determine the temperature of the LPT onset. Confocal fluorescence microscopy of Nile Red stained embryos showed a significant increase in lipid content of the LA treated group compared to the controls. Raman measurements showed that the onset of LPT in LA treated embryos is lower than in untreated ones: −5 °C vs +2 °C. However, these changes in the LPT onset did not affect the survival rates of embryos after cryopreservation. In summary, in vitro culture with LA changes the biophysical characteristics of embryos’ lipidome and is realized in lower LPT onset, but this does not affect embryo survival after cryopreservation.

AB - Lipids significantly affect embryo cryopreservation in some mammalian species depending on the cell lipidome quantity and composition. One of the ways to study the relationship between lipid content and cryotolerance of cells is to study the effect of lipidome modification on laboratory mice. The objective of this research was to study how in vitro culture of mouse embryos with linoleic acid (LA) will affect lipid phase transition (LPT) during cooling and subsequent embryo development after cryopreservation. Embryos obtained in vivo at the 2-cell stage were cultured with 200 μM LA for 46 h up to the morula/blastocyst stage. Thereafter, one portion of embryos was slowly frozen to reveal the effect of LA on survival after cryopreservation, another portion was used to characterize the lipid composition and to determine the temperature of the LPT onset. Confocal fluorescence microscopy of Nile Red stained embryos showed a significant increase in lipid content of the LA treated group compared to the controls. Raman measurements showed that the onset of LPT in LA treated embryos is lower than in untreated ones: −5 °C vs +2 °C. However, these changes in the LPT onset did not affect the survival rates of embryos after cryopreservation. In summary, in vitro culture with LA changes the biophysical characteristics of embryos’ lipidome and is realized in lower LPT onset, but this does not affect embryo survival after cryopreservation.

KW - Cryopreservation

KW - In vitro culture

KW - Intracellular lipids

KW - Linoleic acid

KW - Lipid phase transition

KW - Mouse

KW - Preimplantation embryos

KW - Raman spectroscopy

UR - http://www.scopus.com/inward/record.url?scp=85100033117&partnerID=8YFLogxK

U2 - 10.1016/j.cryobiol.2021.01.014

DO - 10.1016/j.cryobiol.2021.01.014

M3 - Article

C2 - 33485897

AN - SCOPUS:85100033117

VL - 99

SP - 55

EP - 63

JO - Cryobiology

JF - Cryobiology

SN - 0011-2240

ER -