title: Fully automated circulating tumor cell isolation platform with large-volume capacity based on lab-on-a-disc.
abstract: Full automation with high purity for circulating tumor cell (CTC) isolation has been regarded as a key goal to make CTC analysis a "bench-to-bedside" technology. Here, we have developed a novel centrifugal microfluidic platform that can isolate the rare cells from a large volume of whole blood. To isolate CTCs from whole blood, we introduce a disc device having the biggest sample capacity as well as manipulating blood cells for the first time. The fully automated disc platform could handle 5 mL of blood by designing the blood chamber having a triangular obstacle structure (TOS) with lateral direction. To guarantee high purity that enables molecular analysis with the rare cells, CTCs were bound to the microbeads covered with anti-EpCAM to discriminate density between CTCs and blood cells and the CTCs being heavier than blood cells were only settled under a density gradient medium (DGM) layer. To understand the movement of CTCs under centrifugal force, we performed computational fluid dynamics simulation and found that their major trajectories were the boundary walls of the DGM chamber, thereby optimizing the chamber design. After whole blood was inserted into the blood chamber of the disc platform, size- and density-amplified cancer cells were isolated within 78 min, with minimal contamination as much as approximately 12 leukocytes per milliliter. As a model of molecular analysis toward personalized cancer treatment, we performed epidermal growth factor receptor (EGFR) mutation analysis with HCC827 lung cancer cells and the isolated cells were then successfully detected for the mutation by PCR clamping and direct sequencing.
authors.affiliation: Samsung Biomedical Research Institute, Samsung Advanced Institute of Technology, Samsung Electronics Company, Ltd. , 81 Irwon-Ro, Gangnam-Gu, Seoul 135-710, Republic of Korea.
authors.forename: Jong-Myeon
authors.initials: JM
authors.lastname: Park
authors.forename: Minseok S
authors.initials: MS
authors.lastname: Kim
authors.forename: Hui-Sung
authors.initials: HS
authors.lastname: Moon
authors.forename: Chang Eun
authors.initials: CE
authors.lastname: Yoo
authors.forename: Donghyun
authors.initials: D
authors.lastname: Park
authors.forename: Yeon Jeong
authors.initials: YJ
authors.lastname: Kim
authors.forename: Kyung-Yeon
authors.initials: KY
authors.lastname: Han
authors.forename: June-Young
authors.initials: JY
authors.lastname: Lee
authors.forename: Jin Ho
authors.initials: JH
authors.lastname: Oh
authors.forename: Sun Soo
authors.initials: SS
authors.lastname: Kim
authors.forename: Woong-Yang
authors.initials: WY
authors.lastname: Park
authors.forename: Won-Yong
authors.initials: WY
authors.lastname: Lee
authors.forename: Nam
authors.initials: N
authors.lastname: Huh
mesh_terms: D001331:Automation; D001773:Blood Cells; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000295:instrumentation*; D002499:Centrifugation, Density Gradient; D004252:DNA Mutational Analysis; D066246:ErbB Receptors / Q000235:genetics; D006801:Humans; D056656:Lab-On-A-Chip Devices; D044085:Microfluidics; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D016133:Polymerase Chain Reaction; D057285:Precision Medicine
chemical_list: D066246:ErbB Receptors