Intracellular ice formation in mouse zygotes and early morulae vs. cooling rate and temperature-experimental vs. theory

Investigating the factors influencing the characteristics of intracellular ice formation (IIF) is of critical importance for cryopreservation and cryosurgery techniques. However, for the detection of the size of intracellular ice crystals, ∼10nm-0.1μm, there exist serious technical and theoretical difficulties. In this study, a cryomicroscopic method was established to measure the size of intracellular ice crystals in mouse oocytes during their warming processes by investigating the melting point depression of the intracellular ice crystals from extracellular ones. Using the Gibbs-Thomson relation, the size of intracellular ice crystals was calculated and the results range from 4–28 nm, when the molality of the extracellular ethylene glycol and NaCl ranges from 0 to 4m and 0.15 to 0.6m, respectively, and the cooling rate is 100K/min.

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18. Cryoprotectant concentration and cooling rate influence the nucleation temperature and the probability of intracellular ice formation: how and why?

Cryobiology ◽

10.1016/j.cryobiol.2007.10.021 ◽

2007 ◽

Vol 55 (3) ◽

pp. 329

Author(s):

John J. McGrath ◽

Jenny Taubert ◽

Sanket A. Unhale

Keyword(s):

Cooling Rate ◽

Nucleation Temperature ◽

Ice Formation ◽

Intracellular Ice Formation ◽

Intracellular Ice

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Intracellular ice formation in yeast cells vs. cooling rate: Predictions from modeling vs. experimental observations by differential scanning calorimetry

Cryobiology ◽

10.1016/j.cryobiol.2008.11.011 ◽

2009 ◽

Vol 58 (2) ◽

pp. 157-165 ◽

Cited By ~ 32

Author(s):

Shinsuke Seki ◽

F.W. Kleinhans ◽

Peter Mazur

Keyword(s):

Differential Scanning Calorimetry ◽

Cooling Rate ◽

Yeast Cells ◽

Ice Formation ◽

Scanning Calorimetry ◽

Intracellular Ice Formation ◽

Intracellular Ice

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A Study of Intracellular Ice Formation in Jurkat Cells

ASME 2008 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2008-192334 ◽

2008 ◽

Author(s):

Tathagata Acharya ◽

Ram V. Devireddy

Keyword(s):

Cooling Rate ◽

Jurkat Cells ◽

Freezing Rate ◽

Ice Formation ◽

Substantial Evidence ◽

Cooling Rates ◽

Cryoprotective Agents ◽

Intracellular Ice Formation ◽

Intracellular Ice ◽

Phosphate Buffered Saline

The objective of this study was to characterize the IIF behavior of Jurkat cells in isotonic conditions in the absence of any cryoprotective agents. The Jurkat cells were collected from culture and then washed and re-suspended in Dulbecco’s Phosphate Buffered Saline (PBS). The freezing experiments were carried out at defined freezing protocols and at various freezing rates of 5, 20, 30 and 50 °C/min. The results suggest there was no substantial evidence of intracellular ice formation at lower cooling rates of 5, 20 and 30° C/min. The first conspicuous indication of intracellular ice formation (IIF) was observed at a freezing rate of 50 °C/min. At this cooling rate, unlike the usual sudden blackening of cells, the cells suddenly grew and exploded suggesting the formation of intracellular ice, which was reminiscent of a prior observed phenomenon for IIF.

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