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

Tianhang Yang,Songjing Li,Ji Peng,Dayong Gao,Zhiquan Shu,Praveen K. Sekar

doi:10.3390/mi10120832

Tianhang Yang, Songjing Li + Show 4 more

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https://doi.org/10.3390/mi10120832

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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.

Highlights

Freezing is lethal to most living systems, yet cells can endure long periods of storage at low temperatures such as −196 ◦C for centuries [1,2]
During the cooling, thawing, addition and removal of cryoprotective agent (CPA), the changes in the intracellular and intercellular conditions can be lethal to cells [4,5,6,7,8]
Based on Peter Mazur’s “Two-Factors” hypothesis, slow cooling rates will lead to severe cell dehydration as water leaves the cell, whereas fast cooling rates can lead to fatal “ice injury” to the cells due to there being insufficient time for water transport across the membrane leading to intracellular ice formation (IIF) [9]

Summary

Introduction

Freezing is lethal to most living systems, yet cells can endure long periods of storage at low temperatures such as −196 ◦C for centuries [1,2]. A standard cryopreservation protocol involves 7 main steps [3]: pre-processing, CPA addition, cooling to subzero temperatures, storage at low temperature, thawing, dilution and CPA removal, and post-processing. During the cooling, thawing, addition and removal of CPAs, the changes in the intracellular and intercellular conditions can be lethal to cells [4,5,6,7,8]. Osmotic injury and intracellular ice formation injury are both fatal to cells during cryopreservation and they are firmly related to the transmembrane water transportation [10]. The study of transmembrane mass transfer properties of cell membrane will help researchers to optimize cryopreservation protocols to achieve the best cell viability. The most important properties to be determined are cell membrane permeability coefficient to water (hydraulic conductivity, Lp) and cell membrane permeability coefficient to CPAs (Ps)

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