Hyo-Young Jeong

Highly Efficient Assay of Circulating Tumor Cells by Selective Sedimentation with a Density Gradient Medium and Microfiltration from Whole Blood

Isolation of circulating tumor cells (CTCs) by size exclusion can yield poor purity and low recovery rates, due to large variations in size of CTCs, which may overlap with leukocytes and render size-based filtration methods unreliable. This report presents a very sensitive, selective, fast, and novel method for isolation and detection of CTCs. Our assay platform consists of three steps: (i) capturing CTCs with anti-EpCAM conjugated microbeads, (ii) removal of unwanted hematologic cells (e.g., leukocytes, erythrocytes, etc.) by selective sedimentation of CTCs within a density gradient medium, and (iii) simple microfiltration to collect these cells. To demonstrate the efficacy of this assay, MCF-7 breast cancer cells (average diameter, 24 μm) and DMS-79 small cell lung cancer cells (average diameter, 10 μm) were used to model CTCs. We investigated the relative sedimentation rates for various cells and/or particles, such as CTCs conjugated with different types of microbeads, leukocytes, and erythrocytes, in order to maximize differences in the physical properties. We observed that greater than 99% of leukocytes in whole blood were effectively removed at an optimal centrifugal force, due to differences in their sedimentation rates, yielding a much purer sample compared to other filter-based methods. We also investigated not only the effect of filtration conditions on recovery rates and sample purity but also the sensitivity of our assay platform. Our results showed a near perfect recovery rate (~99%) for MCF-7 cells and very high recovery rate (~89%) for DMS-79 cells, with minimal amounts of leukocytes present.

Blood pressure measurement using finger cuff

Many research groups have studied blood pressure measurement in finger artery because of its convenience. But, low accuracy prohibits many hypertension patients from using this device. So, we suggest measurement algorithm that measure systolic and diastolic blood pressure in finger artery. And we also develop calibration method that decreases the error from difference of finger circumference by subjects. We apply our methods for 90 subjects (age from 20 to 49, 55 male, 35 female) to test feasibility of our method by AAMI SP10 standard. The mean difference of our system is plusmn4.7 mmHg for systolic pressure, plusmn4.2 mmHg for systolic pressure. It proved that the feasibility of our method is clinically acceptable (under plusmn5 mmHg)

Micro-slit filter for separation of circulating tumor cells with high recovery and high purity

We present a novel method for separating circulating tumor cells (CTCs) with high recovery and purity at the same time using a micro-slit filter chip and a fully automated fluidic system. Considering white blood cells (WBCs) as big as CTCs are also captured with CTCs during filtration, we amplified the size of CTCs specifically using microbeads (3 μm) coated with anti-Epithelial Cell Adhesion Molecule (anti-EpCAM) to increase the size difference between WBCs and CTCs. The average diameter of MCF-7 cells was increased from 16.5 μm to 23.1 μm. A micro filter chip having an extremely high aspect ratio (AR=3488) rectangular slit was designed to prevent clogging which induces unwanted aggregation, capturing of other small blood cells and consequently decreasing purity. A fully automated fluid control system was implemented for the better reproducibility and the minimization of handling errors. The procedures from blood loading to staining, prior to analysis, were performed automatically. With the optimized condition, separation experiments using 5ml of normal whole blood spiked with 100 MCF-7 cells have demonstrated the reduction of clogging, high recovery (91.1 %) and high purity (52.0 %) at the same time.