Enrichment of circulating tumor cells in tumor-bearing mouse blood by a deterministic lateral displacement microfluidic device

Abstract

Concentration of real tumor cells leaking into blood from cancer was attempted by a deterministic lateral displacement (DLD) microfluidic device. Spiked cultured cell line tumor cells are often used to verify performance of the circulating tumor cells (CTCs) separation methods. Cultured tumor cells are obviously larger than most of hematocytes and considered not to be appropriate as CTC mimics, while there is uncertainty in identifying real CTCs from clinical samples and there is no practical way to examine CTCs leakage into benign cells during the sorting. In this work, blood samples were prepared from tumor-bearing mice whose tumors were induced by implanting cells with GFP expression to living mice. Therefore, CTCs were identified by their fluorescence emission. We succeeded in the enrichment of tumor cells to 0.05% from the blood, in which CTCs were negligibly detected among three million blood cells, and little loss of CTCs was observed.

Size-Based Differentiation of Cancer and Normal Cells by a Particle Size Analyzer Assisted by a Cell-Recognition PC Software

“…8) On the other hand, it has been reported that non-adherent abnormal CTCs can be detected based on the expression of tumor antigens and biophysical deformities. 9,[11][12][13][14][15][16][17][18] The positive expression of CTC protein markers such as cytokeratin -8, -18 and -19, CD44 and the epithelial cell adhesion molecule (EpCAM) is often found in a CTC. 10) To date, the CellSearch system (Veridex, LLC, USA), which is based on immunostaining for CKs + , CD45 -and nucleus + , is the only device that is currently approved by the Food and Drug Administration for CTC detection,.…”

“…[22][23][24] The capture, isolation and enrichment of non-adherent abnormal CTC are based on their difference from hematologic cells in size and biophysical deformities such as such as shape, stiffness and electrical polarizability. [13][14][15][16][17][18][25][26][27][28][29] Many studies have reported that tumor cells are larger in size as compared to hematologic cells and this difference accounts for the…”

Microfluidics cell sample preparation for analysis: Advances in efficient cell enrichment and precise single cell capture

“…Larger size cells will move laterally toward to the side of channel; smaller particles will flow along the original trajectory. [61][62][63][64][65] DLD arrays allow continuous cancer cell enrichment from peripheral blood. However, the cancer cell purity was still very low after enrichment.…”

Open channel deterministic lateral displacement for particle and cell sorting

“…Deterministic lateral displacement (DLD) is a method of particle separation, based on the continuous flow of particles through an array of obstacles that exhibits exceptional resolution in size-based separations. 3 DLD has been used for cell and bioparticle separations such as blood fractionation, 4-8 trypanosome enrichment from blood, 9,10 cancer cell isolation [11][12][13] and CTC cell cluster isolation from whole blood, 14 DNA and exosome separation, 15 and the separation of cells based on parameters other than size, namely shape and deformability, 16,17 and dielectric properties. 18 Early theoretical work by Inglis et al 19 and Davis et al 20 describing the critical size in DLD arrays has been improved upon by studying the effects of post shape.…”