Label-Free Rapid Separation and Enrichment of Bone Marrow-Derived Mesenchymal Stem Cells from a Heterogeneous Cell Mixture Using a Dielectrophoresis Device

Abstract

Bone marrow-derived mesenchymal stem cells (BMSCs) are an important cell resource for stem cell-based therapy, which are generally isolated and enriched by the density-gradient method based on cell size and density after collection of tissue samples. Since this method has limitations with regards to purity and repeatability, development of alternative label-free methods for BMSC separation is desired. In the present study, rapid label-free separation and enrichment of BMSCs from a heterogeneous cell mixture with bone marrow-derived promyelocytes was successfully achieved using a dielectrophoresis (DEP) device comprising saw-shaped electrodes. Upon application of an electric field, HL-60 cells as models of promyelocytes aggregated and floated between the saw-shaped electrodes, while UE7T-13 cells as models of BMSCs were effectively captured on the tips of the saw-shaped electrodes. After washing out the HL-60 cells from the device selectively, the purity of the UE7T-13 cells was increased from 33% to 83.5% within 5 min. Although further experiments and optimization are required, these results show the potential of the DEP device as a label-free rapid cell isolation system yielding high purity for rare and precious cells such as BMSCs.

A Prominent Cell Manipulation Technique in BioMEMS: Dielectrophoresis

“…As a result of the differences in the dielectric polarizability of the particles, they show different DEP mobility in the non-uniform electric field, which enables selective separation and isolation with DEP. DEP-based separation has been applied extensively for the separation of many different types of bioparticles, including stem cells [110,[189][190][191], circulating tumor cells (CTCs) [94,192], blood cells [193][194][195], bacteria [96,[196][197][198], viruses [199,200], and DNA [201,202]. For instance, Song et al reported a DEP-based continuous flow microfluidic device to separate human mesenchymal stem cells (hMSC) from their differentiation progenies (osteoblasts) [191].…”

“…Due to all these prominent advantages, DEP has become an indispensable particle manipulation technique in microchips developed for biomedical applications [91,101,103]. Consequently, DEP has been applied to control the motion of various bioparticles such as cells, including diseased cells [104][105][106], healthy cells [107][108][109], stem cells [98,110], microorganisms [111][112][113][114][115][116], proteins [117][118][119], DNA [120][121][122], and exosomes [123]. DEP, a prominent particle manipulation technique, has been extensively studied by many research groups and scientists in biosensor and POC applications.…”

Current stem cells technologies used in medicine

“…Yoshioka et al separated bone marrow-derived mesenchymal stem cells (BMSCs) from heterogenic cell compound with bone marrow-derived promyelocytes by the DEP method [33]. As the models of BMSCs the human mesenchymal stem cell line (UET-13) were used and human promyelocytic leukaemia cell line (HL-60) as promyelocytes.…”

Electric and Electrochemical Microfluidic Devices for Cell Analysis

“…Recently, the DEP technique was combined with a bipolar electrode system, and the cells were manipulated in a microfluidic device via electrode arrays in the absence of ohmic contacts (Anand et al, 2015). In addition, a DEP device has been used for cell paring by trapping single cells sequentially (Sen et al, 2013b; Yoshimura et al, 2014; Wu et al, 2017), and a microfluidic DEP device was applied for separation of mesenchymal stem cells (MSCs) from a cell mixture according to their dielectric properties (Yoshioka et al, 2018) and controlled cell differentiation (Yoshioka et al, 2016). Moreover, enrichment and detection of target cancer cells were performed by combining DEP manipulation and impedance measurements (Ngoc-Viet and Jen, 2018).…”