title: Novel surface-enhanced Raman scattering-based assays for ultra-sensitive detection of human pluripotent stem cells.
abstract: Human pluripotent stem cells (hPSCs) are a promising cell source for regenerative medicine, but their derivatives need to be rigorously evaluated for residual stem cells to prevent teratoma formation. Here, we report the development of novel surface-enhanced Raman scattering (SERS)-based assays that can detect trace numbers of undifferentiated hPSCs in mixed cell populations in a highly specific, ultra-sensitive, and time-efficient manner. By targeting stem cell surface markers SSEA-5 and TRA-1-60 individually or simultaneously, these SERS assays were able to identify as few as 1 stem cell in 10(6) cells, a sensitivity (0.0001%) which was ∼2000 to 15,000-fold higher than that of flow cytometry assays. Using the SERS assay, we demonstrate that the aggregation of hPSC-based cardiomyocyte differentiation cultures into 3D spheres significantly reduced SSEA-5(+) and TRA-1-60(+) cells compared with parallel 2D cultures. Thus, SERS may provide a powerful new technology for quality control of hPSC-derived products for preclinical and clinical applications.
authors: Division of Pediatric Cardiology, Department of Pediatrics, Emory University School of Medicine and Children's Healthcare of Atlanta, Atlanta, GA, 30322, USA.
authors: Jingjia
authors: J
authors: Han
authors: Wallace H. Coulter Departments of Biomedical Engineering, Emory University and Georgia Institute of Technology, Atlanta, GA, 30322, USA.
authors: Ximei
authors: X
authors: Qian
authors: Division of Pediatric Cardiology, Department of Pediatrics, Emory University School of Medicine and Children's Healthcare of Atlanta, Atlanta, GA, 30322, USA; Wallace H. Coulter Departments of Biomedical Engineering, Emory University and Georgia Institute of Technology, Atlanta, GA, 30322, USA.
authors: Qingling
authors: Q
authors: Wu
authors: Division of Pediatric Cardiology, Department of Pediatrics, Emory University School of Medicine and Children's Healthcare of Atlanta, Atlanta, GA, 30322, USA.
authors: Rajneesh
authors: R
authors: Jha
authors: School of Materials Science and Engineering, University of Science & Technology Beijing, Beijing, China.
authors: Jinshuai
authors: J
authors: Duan
authors: School of Materials Science and Engineering, University of Science & Technology Beijing, Beijing, China.
authors: Zhou
authors: Z
authors: Yang
authors: Division of Pediatric Cardiology, Department of Pediatrics, Emory University School of Medicine and Children's Healthcare of Atlanta, Atlanta, GA, 30322, USA.
authors: Kevin O
authors: KO
authors: Maher
authors: Wallace H. Coulter Departments of Biomedical Engineering, Emory University and Georgia Institute of Technology, Atlanta, GA, 30322, USA; College of Engineering and Applied Sciences, Nanjing University, Nanjing, Jiangsu Province, 210093, China. Electronic address: snie@emory.edu.
authors: Shuming
authors: S
authors: Nie
authors: Division of Pediatric Cardiology, Department of Pediatrics, Emory University School of Medicine and Children's Healthcare of Atlanta, Atlanta, GA, 30322, USA; Wallace H. Coulter Departments of Biomedical Engineering, Emory University and Georgia Institute of Technology, Atlanta, GA, 30322, USA. Electronic address: chunhui.xu@emory.edu.
authors: Chunhui
authors: C
authors: Xu
mesh_terms: D015415:Biomarkers / Q000032:analysis; D058948:Cell Tracking / Q000379:methods*; D002478:Cells, Cultured; D006801:Humans; D039904:Pluripotent Stem Cells / Q000737:chemistry* / Q000145:classification / Q000166:cytology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013059:Spectrum Analysis, Raman / Q000379:methods*; D055746:Stage-Specific Embryonic Antigens / Q000032:analysis*; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D015415:Biomarkers; D055746:Stage-Specific Embryonic Antigens
keywords: Differentiation; Flow cytometry; Human pluripotent stem cell; Nanoparticle; SERS assay