Intracellular ice formation in mouse oocytes subjected to interrupted rapid cooling

Mazur, Péter; Pinn, Irina L.; Kleinhans, F.W.

doi:10.1016/j.cryobiol.2007.06.007

Cited by 31 publications

(37 citation statements)

References 20 publications

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“…We know from previous work [11] that −25°C is some 12 to 15°C above the temperature at which oocytes in 1.5 M EG normally undergo IIF. We know from our recent study [13] that when they are held for 10 min at −25°C, they undergo sufficient dehydration so that most do not flash during the subsequent rapid cooling to −70°C (Ramp 4). However they do turn black upon warming (Ramps 5 and 6).…”

Section: The Linkam Cryostage Freezing Protocols and Rampsmentioning

confidence: 99%

“…These are oocytes that exhibit no darkening during the initial interrupted rapid cooling, but darken during subsequent warming. In our previous study [13], their occurrence was maximal (88%) with a holding time of 10 min at −25°C during the interrupted rapid cool. In the present study with a 12-min hold the percentage was 83%.…”

Section: Relationship Between the Water Contents Of Oocytes After Frementioning

confidence: 99%

“…The oocytes of interest were those that Mazur et al [13] refer to as Class 4; namely, those that show no evidence of darkening during cooling ramps 1-4, but undergo blackening during the hold in Ramp 5. The extent of blackening was visually assigned a score of 0 to 5 based on photomicrographs captured during the hold.…”

Section: Estimating the Degree Of Blackeningmentioning

confidence: 99%

“…The photomicrographs in Fig. 1 in Mazur et al, [13] depict the blackening vs. temperature of three Class 4 oocytes warmed at 10°C/min. The observed degree of blackening in this set and an additional set of nine not published is shown by the solid circles in Fig.…”

mentioning

confidence: 99%

“…Or put differently, to survive freezing, a cell must be cryopreserved in ways that avoid or minimize IIF. In a recent study, Mazur et al [13] investigated intracellular freezing in mouse oocytes subjected to an unorthodox cooling regime referred to as interrupted rapid cooling. In this procedure, cells are cooled rapidly to a temperature above that at which intracellular ice can form, held various times at that temperature, and then cooled rapidly to −70°C or below.…”

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confidence: 99%

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Kinetics and activation energy of recrystallization of intracellular ice in mouse oocytes subjected to interrupted rapid cooling

Seki

Mazur

2008

Cryobiology

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Intracellular ice formation (IIF) is almost invariably lethal. In most cases, it results from the too rapid cooling of cells to below −40°C, but in some cases it is manifested, not during cooling, but during warming when cell water that vitrified during cooling first devitrifies and then recrystallizes during warming. Recently, Mazur et al. [Cryobiol. 55 (2007) 158] dealt with one such case in mouse oocytes. It involved rapidly cooling the oocytes to −25°C, holding them 10 min, rapidly cooling them to −70°C, and warming them slowly until thawed. No IIF occurred during cooling but intracellular freezing, as evidenced by blackening of the cells, became detectable at −56°C during warming and was complete by −46°C. The present study differs in that the oocytes were warmed rapidly from −70°C to temperatures between −65°C and −50°C and held for 3 to 60 min. This permitted us to determine the rate of blackening as function of temperature. That in turn allowed us to calculate the activation energy (Ea) for the blackening process; namely, 27.5 kcal/mole. This translates to about a quadrupling of the blackening rate for every 5° rise in temperature. These data then allowed us to compute the degree of blackening as a function of temperature for oocytes warmed at rates ranging from 10 to 10,000°C/min. A 10-fold increase in warming rate increased the temperature at which a given degree of blackening occurred by 8°C. These findings have significant implications both for cryobiology and cryo-electron microscopy. KeywordsIce; intracellular; recrystallization; activation energy; oocytes; mouse The major cause of death in cells subjected to freezing is the formation of intracellular ice (IIF). Or put differently, to survive freezing, a cell must be cryopreserved in ways that avoid or minimize IIF. In a recent study, Mazur et al. [13] investigated intracellular freezing in mouse oocytes subjected to an unorthodox cooling regime referred to as interrupted rapid cooling. In this procedure, cells are cooled rapidly to a temperature above that at which intracellular ice can form, held various times at that temperature, and then cooled rapidly to −70°C or below. The rationale is the following: Since after the initial rapid cool, the cells are unfrozen, they are by definition supercooled, and because their water is supercooled, it will leave the cells osmotically in response to the lower chemical potential outside. If they are held at this intermediate temperature for sufficient time, they will lose most of their freezable water and *Corresponding author. Fax +1 865 974-8027, E-mail address: E-mail: pmazur@utk.edu (P. Mazur). ✧ Research supported by NIH Grant R01-RR18470Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the prod...

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Section: The Linkam Cryostage Freezing Protocols and Rampsmentioning

confidence: 99%

Section: Relationship Between the Water Contents Of Oocytes After Frementioning

confidence: 99%

Section: Estimating the Degree Of Blackeningmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Kinetics and activation energy of recrystallization of intracellular ice in mouse oocytes subjected to interrupted rapid cooling

Seki

Mazur

2008

Cryobiology

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Cell inactivation and membrane damage after long‐term treatments at sub‐zero temperature in the supercooled and frozen states

Moussa¹,

Dumont²,

Perrier-Cornet³

et al. 2008

Biotech & Bioengineering

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The survival of cells subjected to cooling at sub-zero temperature is of paramount concern in cryobiology. The susceptibility of cells to cryopreservation processes, especially freeze-thawing, stimulated considerable interest in better understanding the mechanisms leading to cell injury and inactivation. In this study, we assessed the viability of cells subjected to cold stress, through long-term supercooling experiments, versus freeze-thawing stress. The viability of Escherichia coli, Saccharomyces cerevisiae, and leukemia cells were assessed over time. Supercooled conditions were maintained for 71 days at -10 degrees C, and for 4 h at -15 degrees C, and -20 degrees C, without additives or emulsification. Results showed that cells could be inactivated by the only action of sub-zero temperature, that is, without any water crystallization. The loss of cell viability upon exposure to sub-zero temperatures is suggested to be caused by exposure to cold shock which induced membrane damage. During holding time in the supercooled state, elevated membrane permeability results in uncontrolled mass transfer to and from the cell maintained at cold conditions and thus leads to a loss of viability. With water crystallization, cells shrink suddenly and thus are exposed to cold osmotic shock, which is suggested to induce abrupt loss of cell viability. During holding time in the frozen state, cells remain suspended in the residual unfrozen fraction of the liquid and are exposed to cold stress that would cause membrane damage and loss of viability over time. However, the severity of such a stress seems to be moderated by the cell type and the increased solute concentration in the unfrozen fraction of the cell suspension.

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An all-37 °C thawing method improves the clinical outcomes of vitrified frozen-thawed embryo transfer: a retrospective study using a case–control matching analysis

Yan

Yao

Yang

et al. 2023

Arch Gynecol Obstet

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Intracellular ice formation in mouse oocytes subjected to interrupted rapid cooling

Cited by 31 publications

References 20 publications

Kinetics and activation energy of recrystallization of intracellular ice in mouse oocytes subjected to interrupted rapid cooling

Kinetics and activation energy of recrystallization of intracellular ice in mouse oocytes subjected to interrupted rapid cooling

Cell inactivation and membrane damage after long‐term treatments at sub‐zero temperature in the supercooled and frozen states

An all-37 °C thawing method improves the clinical outcomes of vitrified frozen-thawed embryo transfer: a retrospective study using a case–control matching analysis

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