title: Label-Free Isolation and mRNA Detection of Circulating Tumor Cells from Patients with Metastatic Lung Cancer for Disease Diagnosis and Monitoring Therapeutic Efficacy.
abstract: We develop an inertial-based microfluidic cell sorter combined with an integrated membrane filter, allowing for size-based, label-free, and high-efficiency separation and enrichment of circulating tumor cells (CTCs) in whole blood. The cell sorter is composed of a double spiral microchannel that hydrodynamically focuses and separates large CTCs from small blood cells. The focused CTCs with the equilibrium position around the midline of microchannel are further captured and enriched by a membrane filter (pore size of 8 μm) attached at the middle outlet. This integrated microfluidic device can process 1 mL of whole blood containing spiked tumor cells (A549, human lung adenocarcinoma epithelial cell line) within 15 min, with the capture efficiency of 74.4% at the concentration as low as tens of A549 cells per mL of whole blood. This microfluidic cell sorter is further adopted for isolation of CTCs from peripheral blood samples of patients with metastatic lung cancer. The immunostaining and CK-19 mRNA detection are applied for identification of captured CTCs, showing that our method can detect 90% of metastatic lung cancer patients before therapy, whereas the commercially used system can only detect 40% of the same patients. We also use the expression of CK-19 mRNA from captured CTCs as an indicator for monitoring the therapeutic efficiency, which correlates well with X-ray computed tomography (CT) assessment of the disease.
authors.affiliation: Beijing Engineering Research Center for BioNanotechnology & CAS Key Laboratory for Biological Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology , Beijing, 100190, China.
authors.forename: Jidong
authors.initials: J
authors.lastname: Wang
authors.affiliation: Beijing Engineering Research Center for BioNanotechnology & CAS Key Laboratory for Biological Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology , Beijing, 100190, China.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Lu
authors.affiliation: Affiliated Hospital of Academy of Military Medical Sciences (307 Hospital) , No. 8 Dongdajie, Beijing, 100071, China.
authors.forename: Chuanhao
authors.initials: C
authors.lastname: Tang
authors.affiliation: Affiliated Hospital of Academy of Military Medical Sciences (307 Hospital) , No. 8 Dongdajie, Beijing, 100071, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Beijing Engineering Research Center for BioNanotechnology & CAS Key Laboratory for Biological Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology , Beijing, 100190, China.
authors.forename: Jiashu
authors.initials: J
authors.lastname: Sun
authors.affiliation: Peking Union Medical College, Chinese Academy of Medical Sciences , Beijing, 100730, China.
authors.forename: Xuan
authors.initials: X
authors.lastname: Mu
authors.affiliation: Peking Union Medical College, Chinese Academy of Medical Sciences , Beijing, 100730, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Urology, Fudan University Shanghai Cancer Center , Shanghai, 200032, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Dai
authors.affiliation: Affiliated Hospital of Academy of Military Medical Sciences (307 Hospital) , No. 8 Dongdajie, Beijing, 100071, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Li
authors.affiliation: Affiliated Hospital of Academy of Military Medical Sciences (307 Hospital) , No. 8 Dongdajie, Beijing, 100071, China.
authors.forename: Hailong
authors.initials: H
authors.lastname: Zhuo
authors.affiliation: Beijing Engineering Research Center for BioNanotechnology & CAS Key Laboratory for Biological Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology , Beijing, 100190, China.
authors.forename: Xingyu
authors.initials: X
authors.lastname: Jiang
mesh_terms: D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology / Q000556:secondary; D046210:Microfluidic Analytical Techniques; D009360:Neoplastic Cells, Circulating / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis*
chemical_list: D012333:RNA, Messenger