A two-factor hypothesis of freezing injury

Mazur, Péter; Leibo, S.P.; Chu, Ernest H. Y.

doi:10.1016/0014-4827(72)90303-5

Cited by 805 publications

(259 citation statements)

References 22 publications

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“…Although the exact mechanisms of cryoinjury have not been fully understood, the inverted U-shaped curve of cell survival after freezing as a function of cooling rate could be well explained by Mazur's "two-factor hypothesis." 4,5 It is assumed that slowly cooled cells are mainly damaged by elevated concentration of intracellular and extracellular solutions (solute effect) resulting from freezing extracellular water while rapidly cooled cells are mainly damaged by intracellular ice formation (IIF). 6 In general, IIF has been recognized as a lethal event to living cells.…”

Section: Introductionmentioning

confidence: 99%

The effect of solution nonideality on modeling transmembrane water transport and diffusion-limited intracellular ice formation during cryopreservation

Zhao¹,

Takamatsu²,

He³

2014

Journal of Applied Physics

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A new model was developed to predict transmembrane water transport and diffusion-limited ice formation in cells during freezing without the ideal-solution assumption that has been used in previous models. The model was applied to predict cell dehydration and intracellular ice formation (IIF) during cryopreservation of mouse oocytes and bovine carotid artery endothelial cells in aqueous sodium chloride (NaCl) solution with glycerol as the cryoprotectant or cryoprotective agent. A comparison of the predictions between the present model and the previously reported models indicated that the ideal-solution assumption results in under-prediction of the amount of intracellular ice at slow cooling rates (<50 K/min). In addition, the lower critical cooling rates for IIF that is lethal to cells predicted by the present model were much lower than those estimated with the ideal-solution assumption. This study represents the first investigation on how accounting for solution nonideality in modeling water transport across the cell membrane could affect the prediction of diffusion-limited ice formation in biological cells during freezing. Future studies are warranted to look at other assumptions alongside nonideality to further develop the model as a useful tool for optimizing the protocol of cell cryopreservation for practical applications. V C 2014 AIP Publishing LLC. [http://dx

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Section: Introductionmentioning

confidence: 99%

The effect of solution nonideality on modeling transmembrane water transport and diffusion-limited intracellular ice formation during cryopreservation

Zhao¹,

Takamatsu²,

He³

2014

Journal of Applied Physics

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“…This alteration is presumed to arise from the solute's ability to change the conformation and stability of the hydrophilic portions of the plasma membrane. Subsequently, when cells are returned to isotonic solutions, the difference in the osmolality within the cell and the external environment triggers the cells to swell beyond their normal isotonic volume and lyse (19). Alternatively upon shrinkage, cells lose portions of membrane, reducing the effective area of the ceil membrane, which (upon return to isotonic conditions) results in cell lysis before their normal volume is recovered (31).…”

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confidence: 99%

Osmotic Effects on Feline Spermatozoa from Normospermic versus Teratospermic Donors

et al. 2000

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“…In biological systems, the two major sources of cryoinjury are ice damage and solution effects, the latter caused by the excessive concentration of intracellular solutes (9,10). Protection against cryoinjury may be achieved by either freezing slowly to induce dehydration by extracellular freezing, thereby minimizing intracellular ice formation, or alternatively by freezing and thawing very rapidly to prevent dehydration while maintaining a small, innocuous, ice crystal size.…”

mentioning

confidence: 99%

Proline: A Novel Cryoprotectant for the Freeze Preservation of Cultured Cells of Zea mays L

Withers¹,

King²

1979

Plant Physiol.

152

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Proline is an effective cryoprotectant for the storage of cultured cells of Zea mays L. in liquid N2. Increased freeze tolerance can be achieved by pregrowth for 3 to 4 days in medium containing proline. Cells cryoprotected with proline have an increased recovery potential when compared with cells cryoprotected with dimethylsulfoxide and glycerol. They also show a reduced postthaw viability loss and greater tolerance of a range of postthaw culture conditions. It is suggested that the mechanism of action of proline may be similar to that in its putative role of conferring protection against natural stresses. It may be protecting the cell against solution effects caused by dehydration during freezing. These findings are discussed in relation to other freeze tolerance enhancing treatments.Freeze preservation in LN3 as a means of genome storage can now be applied to a very limited range of plant tissue cultures. The reasons for the present limitations are not fully understood (1,20,24).In biological systems, the two major sources of cryoinjury are ice damage and solution effects, the latter caused by the excessive concentration of intracellular solutes (9, 10). Protection against cryoinjury may be achieved by either freezing slowly to induce dehydration by extracellular freezing, thereby minimizing intracellular ice formation, or alternatively by freezing and thawing very rapidly to prevent dehydration while maintaining a small, innocuous, ice crystal size. The addition of cryoprotective compounds may reduce cryoinjury in either rapid or slow freezing. In the majority of freeze preservation protocols applied to plant cells, cryoprotectants (usually DMSO and/or glycerol) are used in combination with freezing at a slow rate (1, 24). The high water content of plant cells results in an enhanced susceptibility to cryoinjury. At the level of dehydration required to avoid solution effect stresses, it is still normally necessary to thaw very rapidly to minimize ice recrystallization damage (1,20,24).Recovery of cultures after thawing is dependent upon both a high postthaw viability and further survival through a recovery period preceding renewal of normal metabolic functions and cell division (15,20 In the present study, proline has been used as both a cryoprotectant and pregrowth additive (23) for the freeze preservation of cultured cells of maize. It will be demonstrated that this compound is highly effective in both modes of use and is superior in a number of respects to conventional cryoprotectants. MATERIALS AND METHODSA suspension culture of Zea mays L. (a cell line derived from cultivar B73 [inbred] and kindly supplied by Dr. Ingo Potrykus) was maintained in exponential growth under the conditions described by Potrykus et aL (13). Cells were harvested for freezing 3 or 4 days after subculturing. Additionally, a cell suspension was pregrown for 3 or 4 days in medium supplemented with 5 or 10%1o (w/v) L-proline. Cryoprotectants were prepared using analytical grades of L-proline, glycerol and DMSO, at double th...

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A two-factor hypothesis of freezing injury

Cited by 805 publications

References 22 publications

The effect of solution nonideality on modeling transmembrane water transport and diffusion-limited intracellular ice formation during cryopreservation

The effect of solution nonideality on modeling transmembrane water transport and diffusion-limited intracellular ice formation during cryopreservation

Osmotic Effects on Feline Spermatozoa from Normospermic versus Teratospermic Donors

Proline: A Novel Cryoprotectant for the Freeze Preservation of Cultured Cells of Zea mays L

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