Non-ideal Solution Thermodynamics of Cytoplasm

Ross-Rodriguez, Lisa U.; Elliott, Janet A.W.; McGann, Locksley E.

doi:10.1089/bio.2012.0027

Cited by 13 publications

(21 citation statements)

References 72 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…21,24 To use this equation, the exact contents of the intracellular solution must be known, which brings forward its own problems. In order to solve this issue, studies have used a grouped solute assumption, combining all non-permeating intracellular solutes into one 18,19,25 in a mathematical rigorous way. 22 Ross-Rodriguez et al used the grouped solute assumption to fit non-permeating cell volume data using the osmotic virial equation truncated to the second degree.…”

Section: Introductionmentioning

confidence: 99%

Permeability and Osmotic Parameters of Human Umbilical Vein Endothelial Cells and H9C2 Cells under Non-ideal Thermodynamic Assumptions: A Novel Iterative Fitting Method

2021

Self Cite

View full text Add to dashboard Cite

Cryopreservation is the use of very low subzero temperatures to preserve cells and tissues for later use. This is achieved by controlled cooling in the presence of cryoprotectants that moderate the amount of ice formed. Mathematical modeling of the cryopreservation process is a useful tool to investigate the different variables that affect the results of this process. The changing cell volume during cryopreservation can be modeled using cell membrane water and cryoprotectant permeabilities and the osmotically inactive fraction of the intracellular contents. These three cell-specific parameters have been found previously for different cell types under ideal and dilute assumptions, but biological solutions at subzero temperatures are far from ideal and dilute, especially when cryoprotectants are included. In this work, the osmotic virial equation is used to model the changing cell volume under non-ideal assumptions, and the intracellular environment is described using the grouped solute, which consists of all impermeant intracellular solutes grouped together, leading to two additional cell-specific parameters, the second and third osmotic virial coefficients of the grouped solute. Herein, we present a novel fitting method to efficiently determine these five cell-specific parameters by fitting kinetic cell volume data under non-ideal assumptions and report the results of applying this method to obtain the parameters for two cell types: human umbilical vein endothelial cells and H9C2 rat myoblasts.

show abstract

Section: Introductionmentioning

confidence: 99%

Permeability and Osmotic Parameters of Human Umbilical Vein Endothelial Cells and H9C2 Cells under Non-ideal Thermodynamic Assumptions: A Novel Iterative Fitting Method

2021

Self Cite

View full text Add to dashboard Cite

show abstract

“…The procedural detail provided in this work and further described in a thesis 80 is appropriate to ensure high reproducibility of results, and can be used to optimize the cryopreservation of other types of cells. However, although interrupted cooling protocols provide a means to determine a range of optimal cooling conditions, it must be emphasized that the responses are cell-type specific, since cell responses are governed by cell-specific characteristics such as hydraulic conductivity 58 , membrane solute permeability 59 60 and intracellular solution osmotic behavior 61 . This work provides the understanding and framework necessary to efficiently design protocols in order to achieve remarkable recovery of membrane-intact and functional cells after cryopreservation.…”

Section: Discussionmentioning

confidence: 99%

“…Volume excursions during their addition and removal can be damaging to cell membranes 25 and depending on concentration, cryoprotectants can be toxic which can cause greater damage than osmotic stress 57 . The degree by which cell volumes change depends on: i ) hydraulic conductivity, a membrane characteristic used to describe water diffusion across the cell membrane 58 , ii ) solute permeability, that describes solute diffusion across the cell membrane 59 60 , and iii ) intracellular solution osmotic virial coefficients, used to describe changes in intracellular osmolality as a function of solute concentration 61 . To maximize cryoprotection and minimize toxicity, lower concentrations of cryoprotectants, shorter exposure times and lower temperatures are favorable 22 62 .…”

mentioning

confidence: 99%

Improved Cryopreservation of Human Umbilical Vein Endothelial Cells: A Systematic Approach

Sultani

Marquez‐Curtis

Elliott

et al. 2016

Sci Rep

Self Cite

View full text Add to dashboard Cite

Cryopreservation of human umbilical vein endothelial cells (HUVECs) facilitated their commercial availability for use in vascular biology, tissue engineering and drug delivery research; however, the key variables in HUVEC cryopreservation have not been comprehensively studied. HUVECs are typically cryopreserved by cooling at 1 °C/min in the presence of 10% dimethyl sulfoxide (DMSO). We applied interrupted slow cooling (graded freezing) and interrupted rapid cooling with a hold time (two-step freezing) to identify where in the cooling process cryoinjury to HUVECs occurs. We found that linear cooling at 1 °C/min resulted in higher membrane integrities than linear cooling at 0.2 °C/min or nonlinear two-step freezing. DMSO addition procedures and compositions were also investigated. By combining hydroxyethyl starch with DMSO, HUVEC viability after cryopreservation was improved compared to measured viabilities of commercially available cryopreserved HUVECs and viabilities for HUVEC cryopreservation studies reported in the literature. Furthermore, HUVECs cryopreserved using our improved procedure showed high tube forming capability in a post-thaw angiogenesis assay, a standard indicator of endothelial cell function. As well as presenting superior cryopreservation procedures for HUVECs, the methods developed here can serve as a model to optimize the cryopreservation of other cells.

show abstract

“…[19][20][21] For example, theoretical modeling of cellular osmotic equilibrium, which affects osmotic transport into and out of the cell, requires consideration of non-ideal intracellular thermodynamics due to the crowded cellular environment. 22 Additionally, macromolecular crowding can influence cellular pathology; accelerated amyloid formation has been demonstrated to occur in crowded environments. [23][24][25] The composition of physiological environments often interferes with the analytical methods typically used to characterize virial coefficients.…”

Section: Introductionmentioning

confidence: 99%

Measuring the effects of macromolecular crowding on antibody function with biolayer interferometry

Kim

Yao

Vanam

et al. 2019

mAbs

View full text Add to dashboard Cite

Biotherapeutic proteins are commonly dosed at high concentrations into the blood, which is an inherently complex, crowded solution with substantial protein content. The effects of macromolecular crowding may lead to an appreciable level of non-specific hetero-association in this physiological environment. Therefore, developing a method to characterize the diverse consequences of nonspecific interactions between proteins under such non-ideal, crowded conditions, which deviate substantially from those commonly employed for in vitro characterization, is vital to achieving a more complete picture of antibody function in a biological context. In this study, we investigated non-specific interactions between human serum albumin (HSA) and two monoclonal antibodies (mAbs) by static light scattering and determined these interactions are both ionic strength-dependent and mAbdependent. Using biolayer interferometry (BLI), we assessed the effect of HSA on antigen binding by mAbs, demonstrating that these non-specific interactions have a functional impact on mAb:antigen interactions, particularly at low ionic strength. While this effect is mitigated at physiological ionic strength, our in vitro data support the notion that HSA in the blood may lead to non-specific interactions with mAbs in vivo, with a potential impact on their interactions with antigen. Furthermore, the BLI method offers a high-throughput advantage compared to orthogonal techniques such as analytical ultracentrifugation and is amenable to a greater variety of solution conditions compared to nuclear magnetic resonance spectroscopy. Our study demonstrates that BLI is a viable technology for examining the impact of non-specific interactions on specific biologically relevant interactions, providing a direct method to assess binding events in crowded conditions.

show abstract

Non-ideal Solution Thermodynamics of Cytoplasm

Cited by 13 publications

References 72 publications

Permeability and Osmotic Parameters of Human Umbilical Vein Endothelial Cells and H9C2 Cells under Non-ideal Thermodynamic Assumptions: A Novel Iterative Fitting Method

Permeability and Osmotic Parameters of Human Umbilical Vein Endothelial Cells and H9C2 Cells under Non-ideal Thermodynamic Assumptions: A Novel Iterative Fitting Method

Improved Cryopreservation of Human Umbilical Vein Endothelial Cells: A Systematic Approach

Measuring the effects of macromolecular crowding on antibody function with biolayer interferometry

Contact Info

Product

Resources

About