Spatial Distribution of the State of Water in Frozen Mammalian Cells

Dong, Jinsheng; Malsam, Jason; Bischof, John C.; Hubel, Allison; Aksan, Alptekin

doi:10.1016/j.bpj.2010.08.035

Cited by 50 publications

(49 citation statements)

References 23 publications

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“…Ice 3120 (OH stretching) 26 Amide I 1659 (C = O stretching) 27 Glycerol 851 (C-C stretching) 28 Sorbitol 878 (C-C = O stretching) 29 Glucose 840 (C-C stretching) 30 Sucrose 836 (C-C stretching) 30 Creatine 840 (C-N torsion) 31 suggest that higher solution concentration does not necessarily correlate to higher levels of postthaw recovery. For the same three-component solution compositions used in Figure 2, MATLAB was used to generate scattered interpolant plots that mapped the postthaw recovery measured as a function of solution composition for two of the three components present in solution using 20-25 experimental points (recovery was averaged for vectors with same values for the two components being plotted).…”

Section: Influence Of Osmolarity and Compositionmentioning

confidence: 99%

“…Samples were frozen using a controlled temperature stage described in more detail elsewhere. 26 Raman measurement of frozen MSC cells MSC cells were trypsinized and washed with DPBS solution before being suspended in experimental solutions. Roughly 1 mL of cell suspension was placed on an aluminum sheet, covered with a piece of mica (Ted Pella, Redding, CA) and sealed with Kapton tape (DuPont, Wilmington, DE), to prevent evaporation/sublimation during each experiment.…”

Section: Confocal Raman Systemmentioning

confidence: 99%

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Combinations of Osmolytes, Including Monosaccharides, Disaccharides, and Sugar Alcohols Act in Concert During Cryopreservation to Improve Mesenchymal Stromal Cell Survival

Pollock

Moller-Trane

et al. 2016

Tissue Engineering Part C: Methods

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There is demand for non-dimethyl sulfoxide (DMSO) cryoprotective agents that maintain cell viability without causing poor postthaw function or systemic toxicity. The focus of this investigation involves expanding our understanding of multicomponent osmolyte solutions and their ability to preserve cell viability during freezing. Controlled cooling rate freezing, Raman microscopy, and differential scanning calorimetry (DSC) were utilized to evaluate the differences in recovery and ice crystal formation behavior for solutions containing multiple cryoprotectants, including sugars, sugar alcohols, and small molecule additives. Postthaw recovery of mesenchymal stem cells (MSCs) in solutions containing multiple osmolytes have been shown to be comparable or better than that of MSCs frozen in 10% DMSO at 1°C/min when the solution composition is optimized. Maximum postthaw recovery was observed in these multiple osmolyte solutions with incubation times of up to 2 h before freezing. Raman images demonstrate large ice crystal formation in cryopreserved cells incubated for shorter periods of time (*30 min), suggesting that longer permeation times are needed for these solutions. Recovery was dependent upon the concentration of each component in solution, and was not strongly correlated with osmolarity. It is noteworthy that the postthaw recovery varied significantly with the composition of solutions containing the same three components and this variation exhibited an inverted U-shape behavior, indicating that there may be a ''sweet spot'' for different combinations of osmolytes. Raman images of freezing behavior in different solution compositions were consistent with the observed postthaw recovery. Phase change behavior (solidification patterns and glass-forming tendency) did not differ for solutions with similar osmolarity, but differences in postthaw recovery suggest that biological, not physical, methods of protection are at play. Lastly, molecular substitution of glucose (a monosaccharide) for sucrose (a disaccharide) resulted in a significant drop in recovery. Taken together, the information from these studies increases our understanding of non-DMSO multicomponent cryoprotective solutions and the manner by which they enhance postthaw recovery.

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Section: Influence Of Osmolarity and Compositionmentioning

confidence: 99%

Section: Confocal Raman Systemmentioning

confidence: 99%

Combinations of Osmolytes, Including Monosaccharides, Disaccharides, and Sugar Alcohols Act in Concert During Cryopreservation to Improve Mesenchymal Stromal Cell Survival

Pollock

Moller-Trane

et al. 2016

Tissue Engineering Part C: Methods

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“…Considerations regarding the procurement, optimal storage temperatures (including freeze-thaw cycles) and quality control (QC) tools for tissue and blood samples have recently been reviewed in detail elsewhere (Ahmed 2011;Hubel et al 2014;Shabikhani et al 2014). Biophysics research using methods such as confocal Raman spectroscopy (Dong et al 2010) has been critical for increasing our understanding of nucleation and ice crystal growth within cells and tissues, and has thereby provided an evidence base for biospecimen freezing and low-temperature storage protocols (Hubel et al 2014). The comparative analysis of other biophysical properties of transformed versus nontransformed cells, such as resistance to fluid shear stress , may also be of future relevance to biopreservation technology.…”

Section: The Biospecimen Lifecycle-what Can and Cannot Be Controlledmentioning

confidence: 99%

A critical analysis of cancer biobank practices in relation to biospecimen quality

2015

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There are concerns that a substantial proportion of published research data is not reproducible, which may partially explain the frequent failure to translate pre-clinical results to clinical care. High-quality cancer biospecimens are needed for robust, reproducible research findings, with most researchers obtaining these specimens from cancer biobanks or tumour banks. This review provides an overview of the types of quality control (QC) activities conducted within cancer biobanks that pertain to biospecimen quality and of biospecimen quality reporting tools, including SPREC and BRISQ. We examine how QC assay results and other biospecimen data are communicated from biobanks to researchers, and whether these activities lead to improved biospecimen quality reporting within the literature and/or to improved research outcomes. We also discuss operational factors that limit QC activities within biobanks and evidence gaps requiring further research. In summary, whereas the provision of quality biospecimens is a common aim of cancer biobanks, QC activities remain underreported and are rarely discussed in the literature, compared with other aspects of biobank operations. Further research is required to determine how biobanks can most efficiently optimise biospecimen quality, and how communication between biobanks and researchers can be improved.

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“…Spectroscopic methods are extensively used for studying protein-water-trehalose interactions in mixtures, liposomes, and cells (17,(31)(32)(33)(34). Because of its label-free detection capabilities, Raman spectroscopy has recently been used in studying biological cells (35).…”

Section: Introductionmentioning

confidence: 99%

A Raman Microspectroscopy Study of Water and Trehalose in Spin-Dried Cells

Abazari

Chakraborty

Hand

et al. 2014

Biophysical Journal

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Long-term storage of desiccated nucleated mammalian cells at ambient temperature may be accomplished in a stable glassy state, which can be achieved by removal of water from the biological sample in the presence of glass-forming agents including trehalose. The stability of the glass may be compromised due to a nonuniform distribution of residual water and trehalose within and around the desiccated cells. Thus, quantification of water and trehalose contents at the single-cell level is critical for predicting the glass formation and stability for dry storage. Using Raman microspectroscopy, we estimated the trehalose and residual water contents in the microenvironment of spin-dried cells. Individual cells with or without intracellular trehalose were embedded in a solid thin layer of extracellular trehalose after spin-drying. We found strong evidence suggesting that the residual water was bound at a 2:1 water/trehalose molar ratio in both the extracellular and intracellular milieus. Other than the water associated with trehalose, we did not find any more residual water in the spin-dried sample, intra- or extracellularly. The extracellular trehalose film exhibited characteristics of an amorphous state with a glass transition temperature of ?22°C. The intracellular milieu also dried to levels suitable for glass formation at room temperature. These findings demonstrate a method for quantification of water and trehalose in desiccated specimens using confocal Raman microspectroscopy. This approach has broad use in desiccation studies to carefully investigate the relationship of water and trehalose content and distribution with the tolerance to drying in mammalian cells.

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Spatial Distribution of the State of Water in Frozen Mammalian Cells

Cited by 50 publications

References 23 publications

Combinations of Osmolytes, Including Monosaccharides, Disaccharides, and Sugar Alcohols Act in Concert During Cryopreservation to Improve Mesenchymal Stromal Cell Survival

Combinations of Osmolytes, Including Monosaccharides, Disaccharides, and Sugar Alcohols Act in Concert During Cryopreservation to Improve Mesenchymal Stromal Cell Survival

A critical analysis of cancer biobank practices in relation to biospecimen quality

A Raman Microspectroscopy Study of Water and Trehalose in Spin-Dried Cells

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