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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.

SUBMITTER: Han J

PROVIDER: S-EPMC5391677 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature

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Novel surface-enhanced Raman scattering-based assays for ultra-sensitive detection of human pluripotent stem cells.

Han Jingjia J Qian Ximei X Wu Qingling Q Jha Rajneesh R Duan Jinshuai J Yang Zhou Z Maher Kevin O KO Nie Shuming S Xu Chunhui C

Biomaterials 20160727

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- ...[more]

PMID: 27509304

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Project description:There is a critical need for sensitive rapid point-of-care detection of bacterial infection biomarkers in complex biological fluids with minimal sample preparation, which can improve early-stage diagnosis and prevent several bacterial infections and fatal diseases. A solution-based surface-enhanced Raman scattering (SERS) detection platform has long been sought after for low cost, rapid, and on-site detection of analyte molecules, but current methods still exhibit poor sensitivity. In this study, we have tuned the morphology of the surfactant-free gold nanostars (GNSs) to achieve sharp protruding spikes for maximum SERS enhancement. We have controlled the GNS spike morphologies and optimized SERS performance in the solution phase using para-mercaptobenzoic acid as an SERS probe. To illustrate the potential for point-of-care applications, we have utilized a portable Raman instrument for measurements. For pathogenic agent sensing applications, we demonstrated rapid and sensitive detection of the toxin biomarker pyocyanin (PYO) used as the bacterial biomarker model system. Pyocyanin is a toxic compound produced and secreted by the common water-borne Gram-negative bacterium Pseudomonas aeruginosa, a pathogen known for advanced antibiotic resistance and association with serious diseases such as ventilator-associated pneumonia and cystic fibrosis. The limit of detection (LOD) achieved for PYO was 0.05 nM using sharp branched GNSs. Furthermore, as a proof of strategy, this SERS detection of PYO was performed directly in drinking water, human saliva, and human urine without any sample treatment pre-purification, achieving an LOD of 0.05 nM for drinking water and 0.4 nM for human saliva and urine. This work provides a proof-of-principle demonstration for the high sensitivity detection of the bacterial toxin biomarker with minimal sample preparation: the "mix and detect" detection of the GNS platform is simple, robust, and rapid, taking only 1-2 min for each measurement. Overall, our SERS detection platform shows great potential for point-of-need sensing and point-of-care diagnostics in biological fluids.

Dataset Information

Novel surface-enhanced Raman scattering-based assays for ultra-sensitive detection of human pluripotent stem cells.

Publications

Novel surface-enhanced Raman scattering-based assays for ultra-sensitive detection of human pluripotent stem cells.

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