Abstract
Using the method of pulsed conductometry, the values of electric conductivity of mouse oocytes in 1.0 M solutions of penetrating cryoprotectants have been first determined and the stability of their plasma membranes to the effect of pulsed electric field of increasing intensity has been investigated. For the oocytes, incubated in ethylene glycol (EG), 1,2-propanediol (1,2-PD), 2,3-butanediol (2,3-BD) and dimethyl sulfoxide (DMSO) solutions, the electric conductivity values were (23.6 ± 3.4), (21.3 ± 5.8) and (21.0 ± 2.3) and (23.6 ± 7.1) μS/cm respectively. The values of electric conductivity for mouse oocytes in acetamide (AC) and formamide (FA) solutions were (16.5 ± 6.1) and (16.9 ±± 10.7) μS/cm, respectively. Cryoprotectant solutions, belonging to alcohols (EG, 2,3-BD), as well as DMSO, were shown to have a stabilizing effect on the mouse oocyte plasma membranes under electric field action. The AC and FA solutions caused no stabilizing effect on mouse oocyte membranes, which was manifested in development of irreversible electric breakdown of plasma membranes with increasing electric field intensity. Probl Cryobiol Cryomed 2018; 28(4): 311-321
Highlights
Using the method of pulsed conductometry, the values of electric conductivity of mouse oocytes in 1.0 M solutions of penetrating cryoprotectants have been first determined and the stability of their plasma membranes to the effect of pulsed electric field of increasing intensity has been investigated
We demonstrated the electric conductivity to be an informative cell parameter, characterizing the morphofunctional state of mouse oocytes at different stages of maturity [20, 21, 24], mouse embryos during cleavage [23], as well as at various stages of cryopreservation [22]
The studies were performed in 0.3 M sucrose solution and 1.0 M solutions of penetrating cryoprotectants: ethylene glycol (EG); 1,2-propa-nediol (1,2-PD); 2,3-butanediol (BD); acetamide (AC); formamide (FA); dimethyl sulfoxide (DMSO)
Summary
Using the method of pulsed conductometry, the values of electric conductivity of mouse oocytes in 1.0 M solutions of penetrating cryoprotectants have been first determined and the stability of their plasma membranes to the effect of pulsed electric field of increasing intensity has been investigated. The AC and FA solutions caused no stabilizing effect on mouse oocyte membranes, which was manifested in development of irreversible electric breakdown of plasma membranes with increasing electric field intensity. Оптимизация программ криоконсервирования ооцитов и эмбрионов млекопитающих тесно связана с поиском эффективных криопротекторов, разработкой комбинированных сред замораживания и способов снижения их токсичного влияния на клетки [13, 16, 17, 20, 21]. Решение данной проблемы особенно актуально в связи с разработкой протоколов криоконсервирования ооцитов и эмбрионов методом витрификации, в которых используют высокие концентрации проникающих криопротекторов [16, 20, 21]. В предыдущих работах нами показано, что электрическая проводимость является информативным параметром, характеризующим морфофункциональное состояние ооцитов мыши на разных стадиях зрелости [6, 7, 10], эмбрионов
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