A Spin-Drying Technique for Lyopreservation of Mammalian Cells

  title={A Spin-Drying Technique for Lyopreservation of Mammalian Cells},
  author={Nilay Chakraborty and Anthony Chang and Heidi Y. Elmoazzen and Michael A. Menze and Steven C. Hand and Mehmet Toner},
  journal={Annals of Biomedical Engineering},
Stabilization of cellular material in the presence of glass-forming sugars at ambient temperatures is a viable approach that has many potential advantages over current cryogenic strategies. Experimental evidence indicates the possibility to preserve biomolecules in glassy matrices of low-molecular mobility using “glass-forming” sugars like trehalose at ambient temperatures. However, when cells are desiccated in trehalose solution using passive drying techniques, a glassy skin is formed at the… 

Development of a surface tension mediated technique for dry stabilization of mammalian cells

The technique developed here addresses an important bottleneck of lyoprocessing which requires the fast and uniform desiccation of cellular samples.

A Raman microspectroscopy study of water and trehalose in spin-dried cells.

Cryopreservation of Spin-Dried Mammalian Cells

It was shown that following re-warming to room temperature and re-hydration with a fully complemented cell culture medium, 51% of the spin-dried and vitrified cells survived and demonstrated normal growth characteristics.

The effect of glass-forming sugars on vesicle morphology and water distribution during drying

To explore this hypothesis, a cell is modelled as a lipid vesicle to monitor the water content and membrane deformation during desiccation, and it is found that the concentration dependencies of the diffusivity and viscosity can have profound effects on membrane deformations, which may have significant implications for vesicles damage during the Desiccation process.

Effect of water content on the glass transition temperature of mixtures of sugars, polymers, and penetrating cryoprotectants in physiological buffer

In this study, the effects of moisture content on the glass transition temperature (Tg) of mixtures of sugars, polymers, penetrating cryoprotectants, and phosphate buffered saline (PBS) solutes are investigated.

Effects of Water on Structure and Dynamics of Trehalose Glasses at Low Water Contents and its Relationship to Preservation Outcomes

The viability of T-cells was determined as a function of end water content after microwave-assisted drying in trehalose solutions and molecular modeling revealed that as the water content approached 0.1 gH2O/gdw the matrix formed a large interconnected trehalOSE skeleton with a minimal number of bound water molecules scattered in the bulk.

Dry state preservation of nucleated cells: progress and challenges.

Effective stabilization of nucleated cells for dry storage would be a transformative development in the field of cell-based biosensors and biotechnologic devices, as well as regenerative medicine and other areas in which stem cells have clinical utility when stable long-term storage becomes available without the use of liquid nitrogen and bulky, energetically expensive freezers.

Dry Preservation of Spermatozoa: Considerations for Different Species.

The range of technologies used to dehydrate sperm and the effect of processing and storage conditions on fertilization and embryogenesis using dried sperm are reviewed in the context of reproductive physiology and cellular morphology in different species.

Alternatives to cryopreservation for the short and long-term storage of mammalian cells

There is a growing need to develop scalable GMPcompatible preservation methods that retain not just high cell viability but also clinical efficacy, and this review will explore two alternatives to cryopreservation: cell desiccation for long-term storage of cells and the short-term stored cells under hypothermic (>0°C) conditions.



A role for microwave processing in the dry preservation of mammalian cells

Microwave‐processing successfully facilitated rapid and uniform dehydration of cell‐based samples and improved drying uniformity, thereby overcoming some of the challenges in this field of dry preservation.

Beneficial effect of intracellular trehalose on the membrane integrity of dried mammalian cells.

The results show that a key component of mammalian cells can be preserved in a dried state for weeks under mild conditions and thereby suggest new approaches to preserving mammalian cells.

A simple mechanistic way to increase the survival of Mammalian cells during processing for dry storage.

The present study demonstrates the applicability of this approach and elucidates a simple mechanistic technique to reduce cumulative osmotic stress during processing and demonstrates that cells that were preprocessed to higher trehalose concentrations in step changes prior to drying had higher viability scores at comparable final moisture levels.


  • H. Meryman
  • Biology
    Annals of the New York Academy of Sciences
  • 1960
It has since proved simpler and more effective to freeze the cell suspension by sublimation in the same vacuum chamber in which drying will subsequently proceed, and the freezing, drying, and reconstituting procedures have been extensively varied and investigated.

Survival of Desiccated Mammalian Cells: Beneficial Effects of Isotonic Media

The objective of this study was to examine the effect that the composition of intra- and extracellular trehalose solutions has on the survival of dried 3T3 fibroblasts and found that the post-rehydration membrane integrity and percent cell growth were a function of the residual moisture content of the cells following drying.

Freezing of living cells: mechanisms and implications.

  • P. Mazur
  • Biology
    The American journal of physiology
  • 1984
A new view of the mechanism of slow freezing injury ought to facilitate the development of procedures for the preservation of complex assemblages of cells of biological, medical, and agricultural significance.

Role of trehalose in prevention of giant vesicle adsorption and encapsulated solute leakage in anhydrobiotic preservation.

DLPC vesicles largely avoided adsorption and exhibited high intravesicular solute retention when dried and rehydrated even in the absence of trehalose, despite significant internal structural changes.

Isothermal desiccation and vitrification kinetics of trehalose-dextran solutions.

  • A. AksanM. Toner
  • Materials Science
    Langmuir : the ACS journal of surfaces and colloids
  • 2004
The feasibility of utilizing isothermal, isobaric vitrification as an economical alternative to the preservation technologies currently in use (mainly, cryopreservation and lyophilization) is explored and it was shown that vitrification can be achieved isothermally without crystallization.

Effect of Cell Substrate Interactions on the Desiccation Behavior of Human Fibroblasts

This study investigated the influence of cell substrate interaction during and following desiccation on cell survival in a human fibroblast model and found that samples cultured on biologically active matrices following Desiccation resulted in a significant increase in cell survival.