Chapter 6
Freezing Technology: Control of Freezing, Thawing, and Ice
Nucleation
Peter Kilbride and Julie Meneghel
Abstract
From early dry-ice-based freezers and passive coolers, cryopreservation devices have come a long way. With
increasing interest in the field of cryobiology from new scientific applications, the importance of reliable,
traceable, and reproducible cold chain devices is sure to increase, ensuring more precise cryopreservation
and enabling better post-thaw outcomes, both for the user and for biological samples. As with any
cryopreservation process, it is important to optimize each part of the cold chain for each lab’s biological
samples, cryocontainers used, and logistical restraints. In this chapter we describe how freezing technology
can be used for cryopreservation of cells.
Key words Controlled freezing rate, Ice nucleation, Thawing, Freezing technology
1 Introduction
For successful cryopreservation of most eukaryotic cells, control of
the freezing, cryogenic storage, and thawing procedures is essential
for optimal cell recovery. Recent advances in cellular therapies
employing cryopreservation, often administered directly and
immediately post-thawing, require stringent controls to ensure
that the required level of cell function is achieved without the
need for a “recovery” period of in vitro culture. Such therapies
often require one or several cooling, storage, and warming stages,
with a typical process shown in Fig. 1.
Historically, cryopreservation of cell lines was carried out using
a passive cooling system with samples enclosed in, for example, a
polystyrene box that was then cooled by placing in a suitable
refrigerated environment. More recently low-cost systems such as
a “Mr. Frosty” or “CellCool” designed for use with À80
C refrigerators have become commercially available. These are widely used
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_6, © The Author(s) 2021, Corrected Publication 2021
The original version of this chapter was revised. The correction to this chapter is available at https://doi.org/10.
1007/978-1-0716-0783-1_41
191
Freezing Technology: Control of Freezing, Thawing, and Ice
Nucleation
Peter Kilbride and Julie Meneghel
Abstract
From early dry-ice-based freezers and passive coolers, cryopreservation devices have come a long way. With
increasing interest in the field of cryobiology from new scientific applications, the importance of reliable,
traceable, and reproducible cold chain devices is sure to increase, ensuring more precise cryopreservation
and enabling better post-thaw outcomes, both for the user and for biological samples. As with any
cryopreservation process, it is important to optimize each part of the cold chain for each lab’s biological
samples, cryocontainers used, and logistical restraints. In this chapter we describe how freezing technology
can be used for cryopreservation of cells.
Key words Controlled freezing rate, Ice nucleation, Thawing, Freezing technology
1 Introduction
For successful cryopreservation of most eukaryotic cells, control of
the freezing, cryogenic storage, and thawing procedures is essential
for optimal cell recovery. Recent advances in cellular therapies
employing cryopreservation, often administered directly and
immediately post-thawing, require stringent controls to ensure
that the required level of cell function is achieved without the
need for a “recovery” period of in vitro culture. Such therapies
often require one or several cooling, storage, and warming stages,
with a typical process shown in Fig. 1.
Historically, cryopreservation of cell lines was carried out using
a passive cooling system with samples enclosed in, for example, a
polystyrene box that was then cooled by placing in a suitable
refrigerated environment. More recently low-cost systems such as
a “Mr. Frosty” or “CellCool” designed for use with À80
C refrigerators have become commercially available. These are widely used
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_6, © The Author(s) 2021, Corrected Publication 2021
The original version of this chapter was revised. The correction to this chapter is available at https://doi.org/10.
1007/978-1-0716-0783-1_41
191