Abstract
Effective methods for rapid sorting of cells according to their viability are critical in T cells based therapies to prevent any risk to patients. In this context, we present a novel microfluidic device that continuously separates viable and non-viable T-cells according to their dielectric properties. A dielectrophoresis (DEP) force is generated by an array of castellated microelectrodes embedded into a microfluidic channel with a single inlet and two outlets; cells subjected to positive DEP forces are drawn toward the electrodes array and leave from the top outlet, those subjected to negative DEP forces are repelled away from the electrodes and leave from the bottom outlet. Computational fluid dynamics is used to predict the device separation efficacy, according to the applied alternative current (AC) frequency, at which the cells move from/to a negative/positive DEP region and the ionic strength of the suspension medium. The model is used to support the design of the operational conditions, confirming a separation efficiency, in terms of purity, of 96% under an applied AC frequency of 1.5 × 106 Hz and a flow rate of 20μl/h. This work represents the first example of effective continuous sorting of viable and non-viable human T-cells in a single-inlet microfluidic chip, paving the way for lab-on-a-chip applications at the point of need.
Highlights
Cancer immunotherapy has demonstrated great potential for personalized therapies in both pre-clinical and clinical trials [1]
A dielectrophoresis (DEP) force is generated by an array of castellated microelectrodes embedded into a microfluidic channel with a single inlet and two outlets; cells subjected to positive DEP forces are drawn toward the electrodes array and leave from the top outlet, those subjected to negative DEP forces are repelled away from the electrodes and leave from the bottom outlet
Cell death has been associated with a change in plasma membrane potential (PMP), which www.electrophoresis-journal.com is mainly caused by a change of intracellular potassium and sodium [53,54]
Summary
Cancer immunotherapy has demonstrated great potential for personalized therapies in both pre-clinical and clinical trials [1]. Prior to possible injection into the patient’s body, it is important to efficiently separate viable from non-viable CAR-T cells, as non-viable CAR-T cells can pose severe risks to the patient’s health [7,8] This separation is done under static conditions in a petri dish by using cell viability assays that measure cellular metabolic activity [9]. Fluorescence-activated cell sorting (FACS) that uses flow cytometry can instead allow high-throughput cell separation or sorting [12,13] This method requires sample pre-treatment, as the cells must be stained with a fluorescent dye, and an integrated light source [14].
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