Cryopreservation of Hematopoietic Stem Cells: Emerging Science, Technology and Issues

Fleming, K. K.; Hubel, Allison

doi:10.1159/000104213

Cited by 19 publications

(15 citation statements)

References 75 publications

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“…In fact, the definition of experimental protocols, where the stepwise CPA addition and elution and even the use of automated cell washers are the subjects of research articles recently published. 11,13,39,40 On the other hand, a stepwise CPA addition/removal for the reduction of cell volume variation always implies the increase of the equilibration time. Consequently, to limit longer contact times with a cytotoxic agent that may also be lethal, 41 low temperature equilibration is currently under investigation.…”

Section: Resultsmentioning

confidence: 99%

Rationalizing the equilibration and cooling stages of cryopreservation: The effect of cell size distribution

2011

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A novel model able to quantitatively describe intracellular ice formation (IIF) as a function of temperature in a cell population characterized by a size distribution is proposed for the case when a permeant cryoprotectant agent (CPA) is present. As such, the model represents the ideal extension of the one recently proposed by the authors for interpreting the behavior of a size-distributed population of cells during cryopreservation without CPA. To resemble the experimental procedure actually adopted, the former model is coherently modified for the simulation of the initial equilibration stage (i.e., a one-step CPA permeation kinetics into cells initially under isotonic conditions to be suspended in a water/salt/CPA solution at ambient temperature) and of the subsequent cooling stage at constant rate down to À60 C. Specifically, the size distribution dynamics of a cell population in response to water osmosis, CPA permeation, and IIF is simulated by means of a suitable population balance approach. Water and CPA intracellular transport equations based on the classical Kedem and Katchalsky formalism are coupled to the quantitative description of nucleation and diffusion-limited growth of ice crystals in the framework of a one-dimensional population balance equation. The driving forces of all the physico-chemical phenomena involved are evaluated by taking into account the relevant ternary phase diagram, whereas the viscosity of the cytoplasm solution is properly accounted as a function of the salt and CPA concentrations. After a reasoned choice of the values of model parameters, the behavior of isolated rat hepatocytes suspended in a water/sodium-chloride/glycerol solution is simulated and analyzed at different, practicable (i.e., low) constant cooling rates and CPA equilibration concentrations. It is confirmed that differently sized cells in a single population exhibit different IIF temperatures under the same operative conditions even Correspondence concerning this article should be addressed to A. Cincotti at cincotti@dicm.unica.it or G. Cao at cao@visnu.dicm.unica.it. BIOENGINEERING, FOOD, AND NATURAL PRODUCTSin the presence of CPA. Correspondingly, the probability of IIF results to be a function of the initial size distribution of the cell population. Depending on the specific operative conditions adopted, the size distribution and the osmotic properties of the cell lineage at hand, IIF at À60 C may be lethal for a fraction of the cell population (i.e., larger size classes), or it may not reach a dangerous level for the intermediate size class cells, while it will not even take place for the smaller ones. Finally, even if no comparison with experimental data is provided, an original and physically sound explanation for several, well-known experimental evidences, which appeared in a number of articles available in the literature of cell cryopreservation with CPA, is comprehensively given.

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Section: Resultsmentioning

confidence: 99%

Rationalizing the equilibration and cooling stages of cryopreservation: The effect of cell size distribution

2011

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“…-With safety, energy, health, and the environment issues becoming more pronounced, the utilization of chemicals in preservation processes that are not environmentally friendly or outright harmful is becoming more of an issue. Most of the cryoprotectants are extremely cytotoxic (even mutagenic) at high temperatures (such as the widely used cryoprotectant, dimethyl sulfoxide) and therefore require removal immediately after thawing [27]. This not only adds a critical procedure to be performed before the product can be used but also residual cryoprotectant left in the product causes loss of viability, and function.…”

Section: Current Stabilization Methodsmentioning

confidence: 99%

“…The development of certain technologies, e.g. volumetric rapid cooling, utilization of non-toxic and environmentally friendly stabilization agents, efficient, and safe preservation agent loading, and removal methods [27], without any doubt, will affect the stabilization field in a positive direction. Developments in the spectroscopic and molecular microscopy techniques will help us understand the mechanisms at the molecular level and predict the macroscopic outcome.…”

Section: Future Of Biostabilizationmentioning

confidence: 99%

Protein Stabilization

Ragoonanan

Aksan²

2007

Transfus Med Hemother

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The need for efficient and safe stabilization procedures for biomaterials, therapeutic proteins, and cells is increasing with advances in medicine and pharmaceutics. Stabilization of a protein translates into preservation of the protein structure during storage, in thermodynamic equilibrium with its surroundings. Here, we present a review of the basic thermodynamic principles that govern the protein structural transitions and the interactions of the protein with its surroundings. Specifically, the roles of pressure, temperature, solvent mobility, and solute concentration are discussed. Various methods that can be used to measure protein structure and function, and also the currently available stabilization methods are reviewed. We also introduce a new and promising stabilization method, nanoencapsulation. Similar to the stabilization mechanism of osmolytes, in nanoencapsulation the water activity is altered, affecting the molecular motions of the proteins. There are several distinct advantages of nanoencapsulation, which may also open the way to successful stabilization of biomaterials for bioreactive coatings, biosensors, and biocatalysts.

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“…Cryopreservation, as a means of longterm preservation, is generally required for this reason. 4 The widespread use of HSC treatments has resulted in an increasing demand for cryoprotective techniques and for continuous re-examination of cryoprotective practices. In the past few years, improvements have been made in bone marrow, umbilical cord blood, and peripheral blood cryopreservation techniques.…”

Section: Introductionmentioning

confidence: 99%

Cryopreservation of Peripheral Blood Stem Cells Using a Box-in-Box Cooling Device

Zhou

Kang

Shu

et al. 2009

Biopreservation and Biobanking

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The cooling process is critical for the cryopreservation of human hematopoietic stem cells (HSCs). Currently, programmed freezing methods and uncontrolled cooling methods are in use, both having obvious disadvantages. In this article, a novel device termed Box-in-Box (BIB) was developed and evaluated by in vitro cryopreservation tests in 2 different operation modes ("against-side" mode for Group I (n = 10), and "in-middle" mode for Group II (n = 10), respectively), and compared with an uncontrolled cooling method (Group III (n = 7), Styrofoam boxes) as well as a conventional programmed freezer method (Group IV (n = 10), CryoMed TM 1010, Cryogenic Tech., FL). Recorded temperature profiles of samples cryopreserved with BIB show that a consistent cooling procedure with a rate around -1°C to -3.5°C/min can be achieved during their transfer from room temperature to an -80°C freezer. Statistical analysis of the stem cell population recovery, survival, and colony generation recovery shows that there is no significant difference (P > 0.26) among the methods using the BIB or programmed freezer (Group I, Group II, and Group IV), and their related deviations are smaller than the uncontrolled cooling rate method (Group III). Methods using the BIB (Group I and Group II) generated significantly better cell survival rate (P < 0.01) than the uncontrolled cooling rate method (Group III). The results indicate that the controlled cooling rate methods (BIB or CryoMed PF) are more consistent and reliable for clinical use. Considering the advantages of low cost, durability, and no liquid nitrogen consumption for the cooling process, the BIB can be a good alternative to the programmed freezers for the cryopreservation of HSCs.

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Cryopreservation of Hematopoietic Stem Cells: Emerging Science, Technology and Issues

Cited by 19 publications

References 75 publications

Rationalizing the equilibration and cooling stages of cryopreservation: The effect of cell size distribution

Rationalizing the equilibration and cooling stages of cryopreservation: The effect of cell size distribution

Protein Stabilization

Cryopreservation of Peripheral Blood Stem Cells Using a Box-in-Box Cooling Device

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