Membrane permeability of the human granulocyte to water, dimethyl sulfoxide, glycerol, propylene glycol and ethylene glycol

Abstract

Granulocytes are currently transfused as soon as possible after collection because they rapidly deteriorate after being removed from the body. This short shelf life complicates the logistics of granulocyte collection, banking and safety testing. Cryopreservation has the potential to significantly increase shelf life; however, cryopreservation of granulocytes has proven to be difficult. In this study, we investigate the membrane permeability properties of human granulocytes, with the ultimate goal of using membrane transport modeling to facilitate development of improved cryopreservation methods. We first measured the equilibrium volume of human granulocytes in a range of hypo- and hypertonic solutions and fit the resulting data using a Boyle-van't Hoff model. This yielded an isotonic cell volume of 378 μm3 and an osmotically inactive volume of 165 μm3. To determine the permeability of the granulocyte membrane to water and cryoprotectant (CPA), cells were injected into well-mixed CPA solution while collecting volume measurements using a Coulter Counter. These experiments were performed at temperatures ranging from 4 to 37 °C for exposure to dimethyl sulfoxide, glycerol, ethylene glycol and propylene glycol. The best-fit water permeability was similar in the presence of all of the CPAs, with an average value at 21 °C of 0.18 μm atm−1 min−1. The activation energy for water transport ranged from 41 to 61 kJ/mol. The CPA permeability at 21 °C was 6.4, 1.0, 8.4 and 4.0 μm/min for dimethyl sulfoxide, glycerol, ethylene glycol and propylene glycol, respectively, and the activation energy for CPA transport ranged between 59 and 68 kJ/mol.

Determination of the Membrane Transport Properties of Jurkat Cells with a Microfluidic Device

“…The parameter fitting result of Lp in our study was 0.14 µm•min −1 •atm −1 and Ps (to DMSO) was 0.00034 cm/min in 22 • C. For reference, former results have shown Lp to be 0.38 µm•min −1 •atm −1 and Ps as 0.00049 cm/min for rat basophilic leukemia cell [30], Lp as 0.18 µm•min −1 •atm −1 and Ps as 0.00046 cm/min for human granulocyte [48], and Lp as 0.295 µm•min −1 •atm −1 and Ps as 0.00098 cm/min for human vaginal mucosal macrophage [49]. A considerably convincing result was determined by the parameter-fitting process in this study compared with other results.…”

Comparison of non-ideal solution theories for multi-solute solutions in cryobiology and tabulation of required coefficients

“…Despite the non-ideal thermodynamic nature of the solutions involved, solution models incorporating an ideal dilute assumption are prevalent in cryobiology [8,9,11,12,18,30,31,34,37,39,[61][62][63][64][65][68][69][70]. One commonly-used form of ideal model is to assume that the solution osmolality is equal to the total solute concentration [11,12,18,34,37,61,70].…”

“…One commonly-used form of ideal model is to assume that the solution osmolality is equal to the total solute concentration [11,12,18,34,37,61,70]. This approach can be implemented with concentration expressed in terms of, for example, molality or mole fraction, i.e., respectively…”

Determination of the Membrane Permeability to Water of Human Vaginal Mucosal Immune Cells at Subzero Temperatures Using Differential Scanning Calorimetry

“…There have been many methods proposed and applied by researchers to measure the cell membrane permeability to water, primarily at temperatures above the freezing point. [8][9][10][11][12][13][14][15][16][17][18] However, the membrane permeability at subzero temperatures, for example, in the range of 0°C to -35°C, plays an important role because cell death by solution injury or ice injury occurs in this range. One method to determine the permeability in this range is to measure the L p values at a few suprazero temperatures (e.g., 22°C, 10°C, and 4°C), and then predict the L p at subzero temperatures with the Arrhenius relationship.…”