Single-Cell Isolation of Circulating Tumor Cells from Whole Blood by Lateral Magnetophoretic Microseparation and Microfluidic Dispensing.
This paper introduces a single-cell isolation technology for circulating tumor cells (CTCs) using a microfluidic device (the "SIM-Chip"). The SIM-Chip comprises a lateral magnetophoretic microseparator and a microdispenser as a two-step cascade platform. First, CTCs were enriched from whole blood by the lateral magnetophoretic microseparator based on immunomagnetic nanobeads. Next, the enriched CTCs were electrically identified by single-cell impedance cytometer and isolated as single cells using the microshooter. Using 200 μL of whole blood spiked with 50 MCF7 breast cancer cells, the analysis demonstrated that the single-cell isolation efficiency of the SIM-Chip was 82.4%, and the purity of the isolated MCF7 cells with respect to WBCs was 92.45%. The data also showed that the WBC depletion rate of the SIM-Chip was 2.5 × 10(5) (5.4-log). The recovery rates were around 99.78% for spiked MCF7 cells ranging in number from 10 to 90. The isolated single MCF7 cells were intact and could be used for subsequent downstream genetic assays, such as RT-PCR. Single-cell culture evaluation of the proliferation of MCF7 cells isolated by the SIM-Chip showed that 84.1% of cells at least doubled in 5 days. Consequently, the SIM-Chip could be used for single-cell isolation of rare target cells from whole blood with high purity and recovery without cell damage.
Department of Nano Science and Engineering Center for Nano Manufacturing, Inje University , Gimhae 621-749, Republic of Korea.
Jinho
J
Kim
Department of Nano Science and Engineering Center for Nano Manufacturing, Inje University , Gimhae 621-749, Republic of Korea.
Hyungseok
H
Cho
Department of Nano Science and Engineering Center for Nano Manufacturing, Inje University , Gimhae 621-749, Republic of Korea.
Song-I
SI
Han
Department of Nano Science and Engineering Center for Nano Manufacturing, Inje University , Gimhae 621-749, Republic of Korea.
Ki-Ho
KH
Han
D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D006801:Humans; D061986:MCF-7 Cells; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D059010:Single-Cell Analysis / Q000295:instrumentation; D014407:Tumor Cells, Cultured