Thermodynamics and kinetics of intracellular ice formation during freezing of biological cells

1990 ◽  
Vol 67 (3) ◽  
pp. 1582-1593 ◽  
Author(s):  
Mehmet Toner ◽  
Ernest G. Cravalho ◽  
Marcus Karel
Keyword(s):  
Ice Formation ◽  
Biological Cells ◽  
Thermodynamics And Kinetics ◽  
Intracellular Ice Formation ◽  
Intracellular Ice ◽  
Kinetics Of

Phenomena at the advancing ice–liquid interface: solutes, particles and biological cells

1988 ◽  
Vol 21 (2) ◽  
pp. 229-298 ◽  
Author(s):  
Christoph Körber
Keyword(s):  
Critical Velocity ◽  
Liquid Interface ◽  
Solute Diffusion ◽  
Ice Formation ◽  
Transfer Model ◽  
Square Root ◽  
Biological Cells ◽  
Intracellular Ice Formation ◽  
Intracellular Ice ◽  
Good Agreement

SUMMARYIce formation in aqueous solutions and suspensions involves a number of significant changes and processes in the residual liquid. The resulting effects were described concerning the redistribution of dissolved salts, the behaviour of gaseous solutes and bubble formation, the rejection and entrapment of secondphase particles. This set of conditions is also experienced by biological cells subjected to freezing. The influences of ice formation in that respect and their relevance for cryopreservation were considered as well.A model of transient heat conduction and solute diffusion with a planar ice front, propagating through a system of finite length was found to be in good agreement with measured salt concentration profiles. The spacing of the subsequently developing columnar solidification pattern was of the same order of magnitude as the pertubation wavelengths predicted from the stability criterion. Non-planar solidification of binary salt solutions was described by a pure heat transfer model under the assumption of local thermodynamic equilibrium.The rejection of gaseous solutes and the resulting gas concentration profile ahead of a planar ice front has been estimated by means of a test bubble method, yielding a distribution coefficient of 0·05 for oxygen. The nucleation of gas bubbles has been observed to occur at slightly less than 20-fold supersaturation. The subsequent radial growth of the bubbles obeys a square-root time dependence as expected from a diffusion controlled model until the still expanding bubbles become engulfed by the advancing ice-liquid interface. The maximum bubble radii decrease for increasing ice front velocities.The transition between repulsion and entrapment of spherical latex particles by an advancing planar ice-front has been characterized by a critical value of the velocity of the solidification interface. The critical velocity is inversely proportional to the particle radius as suggested by models assuming an undisturbed ice front. The increase of the critical velocity for increasing thermal gradients shows good agreement with a theoretically predicted square-root type of dependence. Critical velocities have also been measured for yeast and red blood cells.The effect of freezing on biological cells has been analyzed for human lymphocytes and erythrocytes. The reduction of cell volume observed during non-planar freezing agrees reasonably well with shrinkage curves calculated from a water transport model. The probability of intracellular ice formation has been characterized by threshold cooling rates above which the amount of water remaining within the cell is sufficient for crystallization. The cooling rate dependence of viability exhibits largely different maxima for lymphocytes and erythrocytes at about 30 and 4700 K/min, respectively. The decrease of viability above these values has been attributed to the damaging effect of intracellular ice formation.

KrioBlastTM: A Hyper-Fast Cooling and Thawing Scalable Device for Vitrification of Stem and Other Cells in Large Volumes

2012 ◽  
Author(s):  
Vladimir F. Bolyukh ◽  
Igor I. Katkov ◽  
Vsevolod Katkov ◽  
Ilya Yakhnenko
Keyword(s):  
Stem Cells ◽  
Small Sample Size ◽  
Small Sample ◽  
Ice Formation ◽  
Intracellular Ice Formation ◽  
Order Of Magnitude ◽  
Leidenfrost Effect ◽  
Intracellular Ice ◽  
Fast Cooling ◽  
New System

Kinetic (very rapid) vitrification (KVF) is a very promising approach in cryopreservation (CP) of biological materials as it is simple, avoids lethal intracellular ice formation (IIF) and minimizes damaging dehydration effects of extracellular crystallization. Moreover, achieving the ultra-high rates, which would prevent IIF during cooling and devitrification during resuscitation, and achieve KVF for practically any type of cells with one protocol of cooling and re-warming would be the “Holy Grail” of cell cryobiology [3]. However such hyperrapid rates currently require very small sample size which, however, is insufficient for many applications such as stem cells, blood or sperm. As the result, even smallest droplets of 0.25 microliters cannot be vitrified sufficiently fast to avoid the use of potentially toxic external vitrification agents such as DMSO or EG due to the Leidenfrost effect (LFE). In this presentation, we describe an entirely new system for hyperfast cooling of one-two order of magnitude larger samples that we call “KrioBlastTM”, which completely eliminates LFE. We have successfully vitrified up to 4,000 microliters of 15% glycerol solutions, which theoretically corresponds to the critical cooling rate of hundreds of thousands °C/min. We believe that such a system can revolutionize the future cryobiological paradigm.

scholarly journals Innocuous Intracellular Ice Improves Survival of Frozen Cells

2002 ◽  
Vol 11 (6) ◽  
pp. 563-571 ◽  
Author(s):  
Jason P. Acker ◽  
Locksley E. Mcgann
Keyword(s):  
Epithelial Cells ◽  
Cell Survival ◽  
Mdck Cells ◽  
Model Systems ◽  
Ice Formation ◽  
Long Term Storage ◽  
Intracellular Ice Formation ◽  
Intracellular Ice ◽  
Term Storage

Extensive efforts to avoid intracellular ice formation (IIF) during freezing have been central to current methods used for the preservation and long-term storage of cells and tissues. In this study, we examined the effect of intracellular ice formation on the postthaw survival of V-79W fibroblast and MDCK epithelial cells using convection cryomicroscopy and controlled-rate freezing. V-79W and MDCK cells were cultured as single attached cells or as confluent cell monolayers. Postthaw cell survival was assessed using three different indices: the presence of an intact plasma membrane, the ability to reduce alamarBlue, and the capacity to form colonies in culture. Regulating the isothermal nucleation temperature was used to control the incidence of IIF in the model systems. We report that the presence of intracellular ice in confluent monolayers at high subzero temperatures does not adversely affect postthaw cell survival. Further, we show that in the absence of chemical cryoprotectants, the formation of intracellular ice alone improves the postthaw survival of cultured V-79W fibroblast and MDCK epithelial cells. Improved long-term storage of cells and tissues will result by incorporating innocuous intracellular ice formation into current strategies for cryopreservation.

Sign in / Sign up

Export Citation Format

Share Document