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.
Highlights
Mesenchymal stem cells (MSCs), one type of somatic stem cells, possess a self-renewal property and the ability to differentiate into mesodermal lineages, such as chondrocytes, osteocytes, adipocytes [1,2,3], and endodermal [4,5,6] and ectodermal lineages [3,7,8,9]
A DEP device with saw-shaped electrodes was fabricated for the separation and enrichment of Bone marrow-derived mesenchymal stem cells (BMSCs), using a mixture of UE7T-13 and HL-60 cells
Compared to line-shaped electrodes, the saw-shaped electrodes have the advantage of efficient cell enrichment owing to the generation of higher electric field gradient with the same applied voltage
Summary
Mesenchymal stem cells (MSCs), one type of somatic stem cells, possess a self-renewal property and the ability to differentiate into mesodermal lineages, such as chondrocytes, osteocytes, adipocytes [1,2,3], and endodermal [4,5,6] and ectodermal lineages [3,7,8,9]. Long-term cell staining with antibodies may interfere with the clinical use of the cells after separation and is not suitable for cell samples containing blood. The density-gradient method is generally used for cell isolation from the bone marrow, which is based on separation by cell size and density after the collection of tissue samples [13,14,15]. This method does not need cell labelling, it has limitations with regard to purity, repeatability, and long centrifugation time.
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