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

Abstract

The Jurkat cell is an immortalized line of human acute lymphocyte leukemia cells that is widely used in the study of adoptive cell therapy, a novel treatment of several advanced forms of cancer. The ability to transport water and solutes across the cell membrane under different temperatures is an important factor for deciding the specific protocol for cryopreservation of the Jurkat cell. In this study we propose a comprehensive process for determination of membrane transport properties of Jurkat cell. using a novel microfluidic controlled single cell-trapping system. The osmotic behavior of an individual Jurkat cell to water and dimethyl sulfoxide (DMSO), a commonly used cryoprotective agent (CPA), under constant temperature, was recorded under a microscope utilizing the modified microfluidic system. The images of the Jurkat cell under osmotic change were processed to obtain a relationship between cell volume change and time. The experimental results were fitted using a two-parameter transport numeric model to calculate the Jurkat cell membrane permeability to water and DMSO at room temperature (22 °C). This model and the calculated parameters can help scientists optimize the cryopreservation protocol for any cell type with optimal cryoprotective agents and cooling rate for future experiments.

Poroelastic osmoregulation of living cell volume

“…The rise time can be interpreted as the time of pressurization during deswelling, or depressurization during swelling. The consideration of rise time in our simulation is a new attempt to phenomenologically capture the effects of active membrane processes that make the cell membrane a time-dependent barrier to the transport of molecules and water in or out of the cell ( Yamamoto et al, 2014 ; Yang and Hinner, 2015 ; Li et al, 2016 ; Emami et al, 2018 ; Yang et al, 2019 ). Furthermore, cells are not exposed to stepwise, instantaneous osmotic perturbations in vivo ( Haskew-Layton et al, 2008 ; Pilizota and Shaevitz, 2012 ), and in our in vitro experiments, it takes at least a few seconds before cells are fully exposed to the media with different osmolarities despite our efforts to exchange the media immediately.…”

Transcriptional Response in Human Jurkat T Lymphocytes to a near Physiological Hypergravity Environment and to One Common in Routine Cell Culture Protocols

“…Additionally, an external reference data set (GSE94971) [ 43 ] which contained lamina associated domains (LADs) of Jurkat T cells was used to label TADs as lamina associated. The nuclear radius was set at 5 µm in accordance with microscopy results [ 44 ]. The resulting table was then processed by Chrom3D, which generated a set of 5000 structural 3D models that all fulfilled the arrangement conditions derived from the table.…”

Numerical Modeling of Temperature-Dependent Cell Membrane Permeability to Water Based on a Microfluidic System with Dynamic Temperature Control

“…Parameters obtained from previous constant temperature experiments for the same cell can be used as reference settings to make the simulations more reasonable and practical. From previous work, the morphology of Jurkat cells is assumed to be highly spherical and its isotonic cell volume can be set as 810 μm 3 , V b as 49.6% of the isotonic cell volume, 35 L pg as 0.370 μm/atm/min, E a as 7.075 kcal/mol 17 , and T r as 295.15 K (room temperature, 22 °C), and the extracellular environment of the cell is set to be switched immediately from an isotonic to a hypertonic nonpermeating salt-only solution (300 mOsm/L to 375 mOsm/L, 600 mOsm/L and 900 mOsm/L). With the aid of Wolfram Mathematica (Wolfram Research, Champaign, IL), the mathematical solutions of the cell volume changing process under dynamic temperature based on eq 5, with constant temperature changing rates of 1, 5, 10, 20, 30, 45, 60, and 90 K/min from 295.15 K (22 °C) to 310.15 K (37 °C), are calculated and shown in Figure 5 .…”